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Statements

CYLD is phosphorylated. 315 / 315
| 1 1 261 52
sparser
"To understand further the role that CYLD phosphorylation may be playing in ATLL pathogenesis, we analyzed this modification in HTLV-1 transformed T-cell lines, representative of ATLL, as well as in primary human ATLL samples."
32024820
sparser
"We further confirmed the importance of IKKε and TBK1 for CYLD phosphorylation by treatment of cells with MRT67307, followed by immunoprecipitation of cell lysates with the phospho(Ser418)-CYLD-specific antibody (Figure xref )."
29755980
sparser
"CYLD phosphorylation and apoptosis of non-GCB-DLBCL cells were detected."
33827598
sparser
"Transient CYLD phosphorylation thus allows for NF-κB activation to proceed in a stimulus-dependent and transient manner prior to inhibition by the A20 ubiquitin-editing complex."
22435550
sparser
"Whether IKKε is involved in the CYLD phosphorylation in HTLV1-trnasformed T cells is yet to be investigated."
21824392
sparser
"CYLD treated with calf-intestinal phosphatase (CIP) following cotransfection with IKKε was no longer recognized by the phospho-substrate antibody, confirming that the IKKε phospho-substrate antibody specifically recognizes phosphorylated CYLD ( xref )."
19481526
sparser
"These findings not only establish CYLD as a negative regulator of Tax ubiquitination but also suggest a mutual regulatory mechanism in which HTLV1 stimulates CYLD phosphorylation and functional inactivation."
21824392
rlimsp
"Regarding the latter possibility, a recent study suggests that CYLD phosphorylation can also be mediated by the IKK-related kinase IKKε and contributes to IKKε-induced tumorigenesis [38]."
21824392
sparser
"Prostaglandins suppress CYLD and Smad2/3 phosphorylation through a proteasome-dependent mechanism."
31715338
sparser
"We developed experiments and successfully demonstrated BTKis could still promote apoptosis through regulating CYLD phosphorylation in rituximab resistant non-GCB-DLBCL."
33827598
sparser
"Specific CYLD phosphorylation should thus be studied by immunoprecipitation with this antibody followed by CYLD detection via immunoblotting."
29755980
sparser
"In these models, inhibition of TBK1/IKKε using MRT67307 and IKKβ using TPCA showed that both subfamilies could phosphorylate CYLD, with the combination of the two inhibitors having a marked effect in reducing phosphorylation and triggering cell death."
32024820
rlimsp
"We have previously shown that induction of CYLD phosphorylation by TNF-α or mitogens is mediated by the IKK complex [22]."
21824392
sparser
"Phosphorylation of CYLD was blocked in IKKγ-deficient cells and rescued with IKKγ reconstitution (Reiley et al., xref )."
29163038
sparser
"We examined CYLD phosphorylation in MT4 cells after treating them with the SMAC mimetic birinapant (TL32711)."
32024820
sparser
"In turn, IKK phosphorylates and further suppresses the death-signaling function of RIPK1 ( xref ), as well as phosphorylates and inhibits CYLD, a deubiquitinase that dismantles K63-linked poly-ubiquitin chains ( xref )."
30507704
rlimsp
"Phosphorylation of purified CYLD with purified CaMKII for mass spectrometry."
24614225
sparser
"These results indicate that while IKK regulates CYLD under basal conditions, it plays no or a minimal role in mediating further enhanced CYLD phosphorylation that is induced by stimulation of NMDA receptors."
24928390
rlimsp
"Our data further suggest that HTLV1 has developed a mechanism to override the negative-regulatory role of CYLD, and this mechanism involves the induction of CYLD phosphorylation."
21824392
sparser
"Nevertheless, our data suggest that CYLD phosphorylation is a mechanism that mediates constitutive Tax ubiquitination and signaling function in HTLV1-transformed T cells."
21824392
sparser
"A recent study showed that CYLD phosphorylation is elevated in transformed cells and inhibition of this phosphorylation by IKK inhibitors triggers apoptosis, suggesting CYLD as a novel therapeutic target for adult T-cell leukemia [ xref ]."
32512887
rlimsp
"In vitro experiments using purified proteins demonstrated direct phosphorylation and activation of CYLD by the beta catalytic subunit of IKK."
24928390
sparser
"In the same study it was observed that phosphorylation of CYLD in the absence of Ca 2+ also correlated with enhanced DUB activity, albeit at a more modest level [ xref ]."
24928390
sparser
"TBK1 signaling was active in these tumors as measured by elevated pCYLD and pTBK1 levels, and, in consonance with our findings in vitro , CYT387 treatment blocked CYLD phosphorylation and paradoxically increased pTBK1 ( xref )."
24444711
sparser
"Therefore, we used mass spectrometry to determine which residue(s) of CYLD is phosphorylated in vivo ."
19481526
sparser
"The two IKK inhibitors utilized possess distinct inhibitory actions on the IKK complex, providing clues on the mechanism of IKK-mediated phosphorylation of CYLD at the PSD."
24928390
sparser
"In addition, HTLV-1 TAX is found in a complex with CYLD and CYLD is constitutively phosphorylated in HLTV-1 transformed T cells xref ."
32024820
sparser
"To determine if the specific phosphorylation of CYLD is dependent on IKKε and TBK1 as hypothesized earlier, we performed an immunoprecipitation with the phospho(Ser418)-CYLD-specific antibody in IKKε/TBK1 deficient cells (Figure xref )."
29755980
rlimsp
"In response to cellular signals, CYLD becomes transiently phosphorylated and inactivated, thus contributing to the activation of IKK."
21824392
rlimsp
"Since Tax stimulates persistent activation of IKK, we examined the status of CYLD phosphorylation in a large panel of T-cell lines transformed by HTLV1 or Tax."
21824392
sparser
"Because a phosphorylation-defective mutant CYLD protein decreases TNF-α−induced TRAF2 ubiquitylation, it has been hypothesized that CYLD phosphorylation serves as a mechanism for allowing the accumula[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
20347313
sparser
"Our observation that MRT67307 inhibits CYLD phosphorylation in IKKε and TBK1 deficient cells indicates a role for other MRT67307-sensitive kinases in TCR signaling."
29755980
sparser
"Two different IKK inhibitors, IKK16 and tatNEMO, were used to assess possible involvement of IKK in mediating CYLD phosphorylation at the PSD."
24928390
sparser
"Therefore, we sought to determine the effect of CYLD phosphorylation on ubiquitination of known CYLD substrates."
19481526
sparser
"Here, we report that SCF β-TRCP regulates the ubiquitination and degradation of CYLD, a process dependent on prior phosphorylation of CYLD at Ser432/Ser436 by IKK."
24961988
sparser
"In response to mitogens and TNF- α , CYLD is rapidly and transiently phosphorylated, which seems to attenuate its DUB function. xref Interestingly, the phosphorylation of CYLD is mediated by IKK, xref implying that CYLD may not be a major factor involved in feedback control of NF- κ B activation."
19373246
sparser
"It plays a role in numerous signalling pathways, for example it has been shown to phosphorylate CYLD, which in turn activates the NF-κB pathway via deubiquitination of several NF-κB regulator proteins ( xref )."
32324216
sparser
"Taken together these data show that CYLD is phosphorylated upon TCR stimulation and that this phosphorylation is dependent on IKKε and TBK1."
29755980
sparser
"Our results indicated that CYLD phosphorylation is a potential target for pharmacologic modification to improve therapeutic outcomes in non-GCB-DLBCL."
33827598
sparser
"This study provides evidence for a novel function of integrin-ILK and EGF signalling crosstalk in mediating Cyld tyrosine phosphorylation and fast actin-based cytoskeletal rearrangements."
22357970
sparser
"However, it should be noted that, while our work establishes a direct link between phosphorylation of CYLD and its catalytic activity, the earlier studies followed ubiquitination of specific proteins in intact cells."
24928390
sparser
"At the two concentrations of 5 and 10 μM tested, both inhibitors reduced CYLD phosphorylation individually but the combination of these inhibitors was more potent in reducing CYLD phosphorylation (Fig. xref )."
32024820
sparser
"Both IKK inhibitors and overexpression of kinase-inactive TBK/IKK can lower CYLD phosphorylation and trigger cell death ( xref )."
34177595
sparser
"Given the multiple kinases that phosphorylate CYLD and the potential redundancy amongst them as suggested by the inhibitor studies, we explored the possibility of simultaneously inactivating all the kinases."
32024820
sparser
"We provide evidence to show that SMAC mimetics, which are being evaluated as antitumor agents, function similarly to the IKK inhibitors to disrupt CYLD phosphorylation."
32024820
sparser
"The present results indicated that CYLD phosphorylation is a potential clinical therapeutic target for non-GCB-DLBCL, especially for rituximab-resistant relapsed/refractory cases."
33827598
sparser
"But whether BTKis can regulate CYLD phosphorylation to induce apoptosis via down-regulating NFκB in non-GCB-DLBCL remains unclear."
33827598
rlimsp
"These results led us to further examine whether CaMKII promotes phosphorylation of CYLD at the PSD."
24614225
sparser
"The fact that phosphorylation of CYLD can be attenuated by tatNEMO suggests that phosphorylation of CYLD at the PSD requires the regulatory subunit IKKγ, in addition to the catalytic subunits of IKK."
24928390
sparser
"Moreover, CYLD phosphorylation was still inhibited by MRT67307 in IKKε deficient cells."
29755980
sparser
"Our data further suggest that HTLV1 has developed a mechanism to override the negative-regulatory role of CYLD, and this mechanism involves the induction of CYLD phosphorylation."
21824392
rlimsp
"The observation that CYLD can also be phosphorylated in the absence of Ca(2+) in isolated PSDs led us to further explore the regulation of CYLD under basal conditions."
24928390
sparser
"Since Tax stimulates persistent activation of IKK, we examined the status of CYLD phosphorylation in a large panel of T-cell lines transformed by HTLV1 or Tax."
21824392
rlimsp
"Treatment with MRT67307, a small compound inhibitor for IKKε and TBK1, inhibited TCR-induced CYLD phosphorylation."
29755980
sparser
"In these cancers, IKKε-mediated activation of NF-κB signaling is required for transformation, at least in part through phosphorylation of the tumor suppressor CYLD ( xref ; xref ) and TRAF2 ( xref )."
23453972
sparser
"It would be of value to determine whether (and how) CYLD phosphorylation and ubiquitylation can be reversed, or if CYLD can undergo other post-translational modifications that can prevent its phosphor[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
20347313
sparser
"The control peptide had no appreciable effect on the Ca 2+ /calmodulin-dependent phosphorylation of CYLD ( xref , lane 5)."
24614225
sparser
"Both of the IKK sub-families can phosphorylate CYLD, and the combination of MRT67307 and TPCA have a marked effect in reducing CYLD phosphorylation and triggering cell death."
32024820
sparser
"As previously observed in mitogen-stimulated T cells, the upper band represented the phosphorylated CYLD, since it was converted to the basal form upon in vitro phosphatase treatment (Figure xref )."
21824392
sparser
"These results indicate that IKK at the PSD promotes degradation of K63-linked polyubiquitins through phosphorylation of CYLD."
24928390
sparser
"Both CYLD and TRAF2 phosphorylation thus increase ubiquitin-dependent NF-kB signaling."
23333767
rlimsp
"A remarkable finding in the present study is that CYLD is constitutively phosphorylated in a large panel of HTLV1-transformed T cells."
21824392
sparser
"Activation of endogenous CaMKII in isolated PSDs promotes phosphorylation of CYLD."
24614225
sparser
"Since previous basic research indicated that CYLD is a key adaptor that regulates various signaling pathways to modulate diverse physiological processes including immune responses [ xref ], we also want to further investigate the potential effects of Lenalidomide regulating CYLD phosphorylation in non-GCB-DLBCL."
33827598
trips
"The specific Rb (S795) residue was significantly higher phosphorylated in EAC compared to normal esophageal tissue (Wilcoxon paired rank test, p=0.004)."
26058606
sparser
"In addition, CYLD phosphorylation impairs its deubiquitinating function, leading to enhanced RIPK1 K63-linked ubiquitination and survival signal of Adult T-cell leukemia/lymphoma cells [ xref ]."
36202787
sparser
"We next examined the impact on cell survival when CYLD phosphorylation was blocked by the IKK inhibitors following incubation with 5 or 10 μM of the two IKK inhibitors for 24–72 h."
32024820
sparser
"After isolating Myc immune complexes, we determined CYLD phosphorylation by IKKε pSubstrate immunoblot ( xref )."
23453969
sparser
"Importantly, increased CYLD phosphorylation was found in primary ATLL patient samples."
32024820
sparser
"We were prompted to examine the role of CYLD phosphorylation in ATLL lymphomagenesis based on reports that NEMO/IKKβ and IKKε can phosphorylate a cluster of serines between residues 418 and 444 of CYLD resulting in the inhibition of CYLD’s catalytic activity xref , xref ."
32024820
sparser
"Our findings support strategies inhibiting CYLD phosphorylation as novel therapeutic approaches for ATLL patients."
32024820
sparser
"They also observed that Tax-1-expressing cells show an increase in CYLD phosphorylation, a modification that interferes with its enzymatic activity; Tax-induced phosphorylation of CYLD may thus represent a feedback mechanism that hinders host cell defences to viral infection [ xref ]."
31835460
rlimsp
"CYLD phosphorylation can be reversed by endogenous phosphatase activity, indicating another layer of regulation of CYLD phosphorylation at the PSD."
24614225
sparser
"Increase in CYLD phosphorylation is accompanied by an increase in PSD associated DUB activity, as shown by an increase in the degradation rate of added K63-linked tetra-ubiquitin chains (Ub4) ( xref , bottom panels)."
24928390
sparser
"In this study, we aimed to demonstrate that BTKis could induce apoptosis by inhibiting CYLD phosphorylation in non-GCB-DLBCL, including in rituximab-resistant non-GCB-DLBCL."
33827598
sparser
"But in OCI-Ly10R cells, BTK inhibitor PCI-32765 but not rituximab could reduce CYLD phosphorylation and induce apoptosis (Fig.  xref f)."
33827598
rlimsp
"Notably, CYLD facilitates the interaction of EGFR with Cbl-b through its Tyr15 phosphorylation in response to EGF, which leads to fine-tuning of the receptor's ubiquitination and subsequent degradation."
28572092
sparser
"However, these findings suggest that the regulation of CYLD phosphorylation by IKKε contributes to the NF-κB activity necessary for IKKε-mediated cell transformation."
19481526
sparser
"It also remains to be examined whether CYLD phosphorylation also contributes to HTLV1-induced T-cell transformation."
21824392
rlimsp
"Immuno-electron microscopy on hippocampal cultures, using an antibody for CYLD phosphorylated at S-418, revealed that the phosphorylated form of CYLD is present at the PSD under basal conditions."
24928390
sparser
"These reports led us to consider phosphorylation of CYLD by both subfamilies of IKK as the mechanism by which it is suppressed in cancer cells."
32024820
sparser
"Treatment with CYT387 abrogated TBK1/IKKε-induced CYLD phosphorylation in 293T cells, similar to MRT67307 and in contrast to Ruxolitinib ( xref )."
24444711
sparser
"We reasoned that since TAX is known to activate IKK and can associate with CYLD xref , the TAX protein may be sufficient to induce CYLD phosphorylation."
32024820
rlimsp
"Conversely, induction of CYLD phosphorylation may serve as a mechanism by which HTLV1 overrides the inhibitory function of CYLD, leading to the persistent activation of NF-κB."
21824392
sparser
"Combined, these data support the notion that TBK1 exerts partial regulation of TNF-mediated cell death via phosphorylation of CYLD."
34363755
sparser
"When serine in this region is mutated into alanine, CYLD cannot be phosphorylated, and its activity will be significantly enhanced, which will impact on CYLD ubiquitination function, resulting in the generation of apoptosis signal through NFκB signaling pathway [ xref , xref ]."
33827598
sparser
"Conversely, induction of CYLD phosphorylation may serve as a mechanism by which HTLV1 overrides the inhibitory function of CYLD, leading to the persistent activation of NF-κB."
21824392
sparser
"While we could detect immunoprecipitated CYLD in PMA/I stimulated wild-type Jurkat T cells, we did not detect any CYLD upon immunoprecipitation with the phospho(Ser418)-CYLD-specific antibody in the IKKε/TBK1 knock-out cells, indicating that CYLD phosphorylation is indeed dependent on these two kinases."
29755980
sparser
"IKK-mediated CYLD phosphorylation and cellular redox activity."
35109788
sparser
"We also confirmed that the antibody that detects phospho-S418 of CYLD is specific by using it to blot lysates taken from MT4 cells that were transduced with a control shRNA or a CYLD-targeting shRNA to generate CYLD-deficient cells (Supplementary Fig. xref )."
32024820
sparser
"The baseline expression levels of CYLD phosphorylation in non-GCB-DLBCL patients and cell lines were detected by Western Blotting."
33827598
sparser
"Preincubation of PSD fractions with CN21, a peptide inhibitor for both the Ca 2+ -dependent and autonomous forms of CaMKII, had no effect on either the phosphorylation of CYLD or increase in DUB activity ( xref ), indicating that neither of these events are mediated by CaMKII."
24928390
sparser
"Phosphorylated CYLD also reduces resistance to apoptosis by the loss of scaffolding function of RIPK1 necessary to activate NF-κB-mediated transcription."
35109788
sparser
"Therefore, in order to determine which IKK family member(s) can most efficiently phosphorylate CYLD, we cotransfected Myc-CYLD with individual IKK family members fused to GST."
19481526
sparser
"CYLD phosphorylation can be pharmacologically reversed by IKK inhibitors, specifically by TBK1/IKKε and IKKβ inhibitors (MRT67307 and TPCA)."
32024820
sparser
"The synchronously change of CYLD phosphorylation and BTK phosphorylation suggested that CYLD phosphorylation should be regulated through inhibition of BTK phosphorylation, which might regulate tumor cell death in non-GCB-DLBCL."
33827598
rlimsp
"Cellular stimuli induce transient phosphorylation of CYLD, which may contribute to the optimal activation of IKK."
21824392
sparser
"The IKKε/TBK1 inhibitor MRT67307 inhibits CYLD phosphorylation independent of IKKε and TBK1."
29755980
sparser
"BTK inhibitors down-regulated CYLD phosphorylation in non-GCB-DLBCL."
33827598
sparser
"These observations strongly suggest that in ATLL cells, either viral encoded oncogenic proteins or mutations sustain the early cell death checkpoint by driving multiple IKKs to phosphorylate and suppress CYLD in order to prevent RIPK1 from becoming a death-signaling molecule (Fig. xref )."
32024820
sparser
"Interestingly, in contrast to the situation in PSD fractions, addition of purified IKKβ was sufficient to phosphorylate purified CYLD."
24928390
sparser
"Here we show that specific phosphorylation of CYLD upon immunoprecipitation is already completely inhibited at 2 μM MRT67307 (Figure xref , left panel), while 10 μM MRT67307 is necessary to completely prevent the phosphorylation band in the total lysate (Figure xref , right panel)."
29755980
rlimsp
"A CaMKII inhibitor CN21 had no effect on CYLD phosphorylation in the absence of Ca(2+), but two different IKK inhibitors, IKK16 and tatNEMO, inhibited its phosphorylation."
24928390
rlimsp
"Interestingly, CYLD undergoes constitutive phosphorylation in HTLV1-transformed T cells, a mechanism known to inactivate the catalytic activity of CYLD."
21824392
sparser
"This suggests that both subfamilies of IKK can phosphorylate CYLD and that there is redundancy amongst the kinases in TAX-expressing cells."
32024820
sparser
"IKKe also phosphorylates and inactivates the tumor suppressor CYLD, preventing CYLD from deubiquitinating specific substrates in the NF-kB signaling pathway."
23333767
sparser
"Jessica Hutti presented evidence that IKKε can phosphorylate CYLD at Ser418, and that CYLD phosphorylation is required for IKKε-induced transformation of cells in culture."
19287453
sparser
"Several DUB enzymes are regulated by serine phosphorylation, including TNFAIP3 (A20) whose DUB activity is increased upon phosphorylation [57] , In contrast, serine phosphorylation of CYLD and OTUB1 [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
23333852
sparser
"In contrast, CYLD is phosphorylated inefficiently in cells transfected with IKKα and IKKβ, even when greater than 5× more kinase is present ( xref )."
19481526
sparser
"ATLL cells overexpressing a kinase-inactive TBK1 (TBK1-K38A) demonstrate lower CYLD phosphorylation and subsequently reduced proliferation."
32024820
sparser
"Chemical inhibition of TBK1/IKKε with MRT also abolished CYLD phosphorylation in response to TNF in WT and NEMO-null Jurkats ( xref ) and increased cell death beyond the impact of NEMO deficiency ( xref , xref )."
34363755
sparser
"Together, these data indicate that CaMKII activation promotes phosphorylation of CYLD at the PSD."
24614225
sparser
"In both of these, there was constitutive phosphorylation of CYLD, accompanied by phosphorylation of IKKα/β and TBK1/IKKε, indicative of active kinases."
32024820
sparser
"These results further suggested that CYLD phosphorylation is an essential mediator in this BTKis downstream pathway to promote cell death in non-GCB-DLBCL."
33827598
sparser
"We speculate that, with insufficient amounts of OTULIN, phosphorylated CYLD primed for activity towards K63-Ub is quenched by M1-Ub."
35587511
sparser
"Interestingly, CYLD undergoes constitutive phosphorylation in HTLV1-transformed T cells, a mechanism known to inactivate the catalytic activity of CYLD."
21824392
sparser
"The role of IKK in promoting CYLD phosphorylation at the PSD under both basal and excitatory conditions was examined using a cell permeable IKK inhibitor, IKK16."
24928390
sparser
"Tyrosine phosphorylated CYLD recruits the ubiquitin ligase Cbl-b, facilitating close proximity of Cbl-b to EGFR that results in ubiquitination and proteasomal degradation of EGFR ( xref )."
32883697
sparser
"When the CYLD immunoprecipitate was incubated in an in vitro kinase assay with WT IKKε, strong phosphorylation of CYLD was observed ( xref )."
19481526
sparser
"However, cotransfection of IKKε with CYLD resulted in the stabilization of NEMO ubiquitination, suggesting that phosphorylation of CYLD decreases its deubiquitinase activity."
19481526
rlimsp
"Thus, CYLD undergoes constitutive phosphorylation in HTLV1-transformed T cells."
21824392
sparser
"Our results indicated that CYLD phosphorylation is a potential pharmacologic target which can sensitize rituximab-resistant non-GCB-DLBCL to BTK inhibitors."
33827598
sparser
"However, within HTLV-transformed cells, CYLD is constitutively phosphorylated which inhibits its deubiquitinase activity and, therefore, Tax association with IKK is maintained (Wu et al., xref )."
23226145
sparser
"Our observations suggest that in cells transformed by HTLV-1, TAX induces the phosphorylation of CYLD to keep it inactive in order to prevent RIPK1 from inducing cell death."
32024820
rlimsp
"The catalytic activity of CYLD is negatively regulated by its phosphorylation [22]."
21824392
sparser
"Recently, IκB kinase epsilon (IKKϵ), an oncoprotein overexpressed in 30% of breast cancers (120), which plays a critical role in interferon signalling pathways, has been shown to phosphorylate and inactivate CYLD (121)."
20073038
sparser
"Prior work discovered that rapid and transient phosphorylation of CYLD is necessary for the ubiquitination of TRAF2 and activation of JNK (Lork et al., 2018)."
34966378
sparser
"IKK inactivates the deubiquitination activity of CYLD by activating the phosphorylation of CYLD, thus promoting the ubiquitination of Nrf2 and further aggravating oxidative stress injury of the kidney in ORN."
34711178
sparser
"Phosphorylation of CYLD serves as a mechanism that inactivates its DUB activity."
23797609
sparser
"Although TNF was previously shown to trigger CYLD phosphorylation in Jurkat, HeLa, or HEK 293 cells, leading to the appearance of a slower migrating form of CYLD (Reiley et al., xref ), stimulation of our Jurkat T cells with TNF did not lead to a band shift or detection of a band with the phospho(Ser418)-specific antibody, suggesting either no phosphorylation or phosphorylation at other sites."
29755980
rlimsp
"In response to cellular stimuli, CYLD undergoes rapid and transient phosphorylation, which is required for signal-induced TRAF2 ubiquitination and activation of downstream signaling events."
15870263
sparser
"These findings indicated that phosphorylation of the residues in CYLD is critical for innate immune response."
34966378
sparser
"Phosphorylation of CYLD in the NF-κB Pathway."
21480003
sparser
"Transfection of a TAX-encoding plasmid into HEK293 EBNA cells confirmed that TAX by itself is sufficient to induce CYLD phosphorylation (Fig. xref )."
32024820
sparser
"Regarding the latter possibility, a recent study suggests that CYLD phosphorylation can also be mediated by the IKK-related kinase IKKε and contributes to IKKε-induced tumorigenesis [ xref ]."
21824392
sparser
"Phosphorylation of CYLD was assessed through changes in its electrophoretic mobility in Western Immunoblots."
24614225
sparser
"CYLD and A20 were phosphorylated during necrosome signaling (Fig.  xref )."
29789521
sparser
"Fibroblasts derived from patients with TBK1 deficiency display heightened sensitivity to necroptosis that results from the lack of inhibitory phosphorylation on CYLD, which increases its deubiquitylation activity towards RIPK1."
35523963
sparser
"To examine whether TBK1/IKKi dual inhibitors suppress CYLD phosphorylation, SCC-9 cells were treated with different TBK1/IKKi dual inhibitors for 1 hr, and cells were collected for western blot."
24150799
sparser
"A remarkable finding in the present study is that CYLD is constitutively phosphorylated in a large panel of HTLV1-transformed T cells."
21824392
sparser
"Experiments with an antibody specific to CYLD phosphorylated at S-418 (p-CYLD) show that incubation of PSD fractions in the presence of ATP in EGTA-containing medium induces phosphorylation of CYLD ( xref , top panels), confirming previous results by mass spectrometry [ xref ]."
24928390
sparser
"As the phosphomimetic CYLD mutant is competent in TRAF2 binding, phosphorylation of CYLD may not interfere with its association with TRAF2, but probably affects the catalytic activity of CYLD."
19373246
sparser
"Similarly, CYLD is also known to inhibit NFκB signaling and therefore, inhibitory phosphorylation of CYLD will also serve to enhance expression of prosurvival genes."
32024820
sparser
"Activation of CaMKII in isolated PSDs promotes phosphorylation of CYLD on the same residues and also enhances endogenous deubiquitinase activity specific for K63-linked polyubiquitins."
24614225
sparser
"Phosphorylation of CYLD has been reported to inhibit its deubiquitinase (DUB) activity xref – xref ."
29789521
sparser
"As further validation, we assessed whether CYLD phosphorylation modulates NF-κB activity in the context of transformation."
19481526
rlimsp
"CYLD is constitutively phosphorylated in HTLV1-transformed T cells."
21824392
sparser
"Cellular stimuli induce transient phosphorylation of CYLD, which may contribute to the optimal activation of IKK."
21824392
sparser
"Specifically, we failed to detect phosphorylation of CYLD in U2OS shUb-Ub(K63R) cells as assessed by a pCYLD-specific antibody."
23453969
rlimsp
"Phosphorylation of expressed CYLD using PSD fraction as source of kinase for mass spectrometry."
24614225
sparser
"However, by immunoprecipitation with the phospho(Ser418)-CYLD-specific antibody followed by CYLD detection via immunoblotting and detection with anti-CYLD, we were able to detect specific CYLD phosphorylation, and could show that this was indeed dependent on IKKε and TBK1, confirming our initial hypothesis."
29755980
sparser
"TBK1-K38A expression blocked CYLD phosphorylation and inhibited the proliferation of MT4 cells."
32024820
rlimsp
"However, we found that in HEK293 cells, Tax was insufficient for the induction of CYLD phosphorylation (data not shown)."
21824392
sparser
"CYLD was immunoprecipitated using an anti-Myc antibody and blotted using the anti-IKKε phospho-substrate antibody. xref shows that CYLD was efficiently phosphorylated in cells transfected with IKKε."
19481526
rlimsp
"Nevertheless, our data suggest that CYLD phosphorylation is a mechanism that mediates constitutive Tax ubiquitination and signaling function in HTLV1-transformed T cells."
21824392
rlimsp
"Regulation of the deubiquitinating enzyme CYLD by IkappaB kinase gamma-dependent phosphorylation."
15870263
sparser
"Whether CYLD phosphorylation is also a regulator for non-GCB-DLBCL apoptosis needs further research."
33827598
sparser
"CYLD is constitutively phosphorylated in HTLV1-transformed T cells."
21824392
sparser
"Our finding appears to be in contradiction with previous reports, indicating that IKK-mediated phosphorylation of CYLD promotes ubiquitination of TRAF2 in HeLa and HEK cells [ xref ], [ xref ] and of IKKγ in HEK cells [ xref ]."
24928390
sparser
"Furthermore, Tax and CYLD appear to mutually regulate, since CYLD is constitutively phosphorylated in HTLV1-transformed T cells."
21824392
sparser
"Both CYLD and TRAF2 phosphorylation thus increase ubiquitin-dependent NF-κB signaling."
23333767
sparser
"Interestingly, TBK1 or IKKε but not RIG-I specifically precipitates two species of CYLD in size, suggesting phosphorylation of CYLD by these kinases."
18703349
sparser
"We demonstrated that BTKis could down-regulate CYLD phosphorylation and induce CYLD dependent apoptosis in non-GCB-DLBCL via inactivating NFκB pathway."
33827598
rlimsp
"In search for specific markers, paraffin-embedded tissue samples from 25 SCC and 64 KA were arranged in a tissue microarray (TMA) and stained for immunologic cell-markers CD3, CD20 and CD68 as well as for proteins considered of relevance in tumorgenesis, namely NF kappaB/p65, I kappaB-alpha, STAT3, p53, TRAP-1, pRB, phosphorylated pRb, Cyld, p21, p16(INK4), Survivin, Bcl-xL, Caspase 3, Bak, FLK-1/VEGF-r2 and Ki-67."
19165861
rlimsp
"Together, these data indicate that CaMKII activation promotes phosphorylation of CYLD at the PSD."
24614225
sparser
"Additionally, in primary adult T-cell leukemia/lymphoma (ATLL) samples and cell lines, increased IKK-induced CYLD phosphorylation was observed."
34177595
sparser
"Activation of IKK in isolated PSDs also promoted phosphorylation of CYLD and an increase in endogenous deubiquitinase activity for K63-linked polyubiquitins."
24928390
sparser
"We have validated this method through the identification of CYLD as a novel IKKε substrate and have shown that CYLD phosphorylation is necessary for full IKKε-mediated anchorage-independent growth."
19481526
sparser
"While there are limitations with the pharmacological and dominant-negative approaches used in this study, nonetheless the observations strongly suggest that phosphorylation of CYLD is a mechanism used by ATLL cells to suppress this tumor suppressor and maintain tumor cell survival."
32024820
sparser
"To determine the effect of the K30 and K401 mutants on IKKε kinase activity, we assessed the ability of WT and mutant IKKε to phosphorylate CYLD in vivo ."
23453969
rlimsp
"Phosphorylation of CYLD was assessed through changes in its electrophoretic mobility in Western Immunoblots."
24614225
sparser
"At the functional level, CYLD phosphorylation was previously shown to prevent CYLD-mediated TRAF2 deubiquitination and to promote TNF-induced gene expression (Reiley et al., xref )."
29755980
sparser
"Birinapant reduced CYLD phosphorylation as well as TBK1/IKKε and IKKα/β phosphorylation (Fig. xref ) and induced cell death in the MT4 cells, which was dependent on CYLD (Fig. xref )."
32024820
sparser
"Finally, we examined the phosphorylation status of CYLD in lysates of human ATLL cryo-preserved samples from which we were able to obtain sufficient protein to resolve by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for CYLD phosphorylation."
32024820
sparser
"Based on these prior reports and the data obtained thus far with the MT4 cells, we hypothesized that TAX activation of IKK family of kinases induces the phosphorylation of CYLD to suppress its deubiquitinating activity."
32024820
sparser
"CaMKII-mediated phosphorylation of CYLD enhances its deubiquitinase activity as shown in in vitro experiments using purified proteins."
24614225
rlimsp
"Effect of CaMKII-mediated phosphorylation on deubiquitinase activity of purified CYLD."
24614225
sparser
"Initial evidence for post-translational regulation of CYLD came from the finding that CYLD undergoes transient phosphorylation in cells stimulated with NF- κ B inducers, such as TNF- α , mitogens, and LPS. xref The phosphorylation of CYLD occurs in a serine cluster located just upstream of its TRAF2-binding site (amino acid 418–444)."
19373246
sparser
"Interestingly, phosphorylation of CYLD has been shown to inhibit its DUB activity xref – xref ."
29789521
rlimsp
"These findings not only establish CYLD as a negative regulator of Tax ubiquitination but also suggest a mutual regulatory mechanism in which HTLV1 stimulates CYLD phosphorylation and functional inactivation."
21824392
sparser
"Recently, IjB kinase epsilon (IKKe), an oncoprotein overexpressed in 30% of breast cancers (120), which plays a critical role in interferon signalling pathways, has been shown to phosphorylate and inactivate CYLD (121) ."
20073038
rlimsp
"NMDA treatment further promoted phosphorylation of CYLD at the PSD, but IKK16 failed to block the NMDA-induced effect."
24928390
rlimsp
"CYLD has constitutive DUB activity, but its activity can be rapidly inactivated via its phosphorylation in response to NF-κB stimuli [22]."
21824392
sparser
"CYLD phosphorylation can be reversed by endogenous phosphatase activity, indicating another layer of regulation of CYLD phosphorylation at the PSD."
24614225
sparser
"IKKɛ also phosphorylates and inactivates the tumor suppressor CYLD, preventing CYLD from deubiquitinating specific substrates in the NF-κB signaling pathway."
23333767
rlimsp
"Constitutive phosphorylation of CYLD in HTLV1-transformed T cells."
21824392
sparser
"Other work shows that CYLD can be phosphorylated upon overexpression of the IKK-related kinase IKKε, facilitating IKKε-driven cellular transformation (Hutti et al., xref )."
29755980
sparser
"However, we found that in HEK293 cells, Tax was insufficient for the induction of CYLD phosphorylation (data not shown)."
21824392
sparser
"Prior work discovered that rapid and transient phosphorylation of CYLD is necessary for the ubiquitination of TRAF2 and activation of JNK ( xref )."
34966378
sparser
"NMDA treatment further promoted phosphorylation of CYLD at the PSD, but IKK16 failed to block the NMDA-induced effect."
24928390
sparser
"Here, IκB kinase (IKK) was hypothesized to inactivate the deubiquitination activity of cylindromatosis (CYLD) by activating the phosphorylation of CYLD, thus promoting the ubiquitination of NF-E2-related factor 2 (Nrf2) and further aggravating oxidative stress injury of the kidney in ORN."
34711178
sparser
"In this study, we put forward a hypothesis of BTK inhibitors (BTKis) could regulate CYLD phosphorylation to promote apoptosis through down-regulating NFκB signaling pathway in non-GCB-DLBCL and we did experiments in vivo and in vitro to demonstrate this hypothesis."
33827598
rlimsp
"Phosphorylation of CYLD under basal conditions was inhibited by IKK16."
24928390
sparser
"To investigate further if IKK kinases are involved in phosphorylating CYLD in MT4 cells, we examined the effect of MRT67307 (TBK1/IKKɛ inhibitor) and TPCA (IKKβ inhibitor)."
32024820
sparser
"These observations indicate that endogenous IKK activity in isolated PSDs mediates CYLD phosphorylation in the absence of Ca 2+ ."
24928390
sparser
"In future studies, it would be beneficial to identify CYLD’s substrates and compare factors involved in orchestrating downstream effects of CYLD phosphorylation between neuronal and non-neuronal cells."
24928390
sparser
"Since a phospho-mimetic CYLD mutant is defective in inhibiting Tax ubiquitination, the CYLD phosphorylation may contribute to the chronic activation of IKK and NF-κB in HTLV1-transformed T cells."
21824392
sparser
"Therefore, one possible explanation for the discrepancy may be that the effect of CYLD on the ubiquitination of certain proteins in intact cells is indirect, mediated by factors downstream from CYLD phosphorylation."
24928390
sparser
"Phosphorylated CYLD elevated in non-GCB DLBCL."
33827598
sparser
"However, CYLD does not have a compensatory role for A20, because of the attribution of TLR and TNFR signals to the inactivation of CYLD in stimulated cells by CYLD phosphorylation [ xref ]."
33138315
sparser
"These findings suggest that CYLD serves as a novel target of IKK and that the site-specific phosphorylation of CYLD regulates its signaling function."
15870263
sparser
"The exact mechanism of CYLD phosphorylation remains to be investigated since two reports, [ xref ] and [ xref ], suggest that phosphorylation either inhibits or activates CYLD, respectively."
27913667
sparser
"Western blotting showed that BTKis inhibited the phosphorylation of both CYLD and BTK in these two cell lines (Fig.  xref a–d)."
33827598
sparser
"A similar mechanism is thought to occur in breast carcinogenesis where IKKε phosphorylates and inactivates CYLD ( xref )."
22435550
sparser
"As IKK-mediated phosphorylation of CYLD has been reported to negatively regulate CYLD enzymatic activity [ xref ], we further explored whether IKK is also involved in regulating the degradation of CYLD by SCF β-TRCP ."
24961988
sparser
"BTK inhibitors can enhance the apoptosis of non-GCB-DLBCL cells induced by rituximab through down-regulating the phosphorylation of CYLD."
33827598
rlimsp
"These findings suggest that CYLD serves as a novel target of IKK and that the site-specific phosphorylation of CYLD regulates its signaling function."
15870263
sparser
"Treatment with MRT67307, a small compound inhibitor for IKKε and TBK1, inhibited TCR-induced CYLD phosphorylation."
29755980
sparser
"Phosphorylation of ubiquitin carboxyl-terminal hydrolase CYLD, a tumor suppressor, and an important player in the host defense mechanism against bacterial infection, at Ser418 by the inhibitor of nuclear factor-κB kinase (IKKγ), impairs its DUB activity."
32512887
sparser
"Interestingly, TBK1 or IKKε specifically causes CYLD to shift to higher molecular bands, suggesting phosphorylation of CYLD by these kinases."
32215003
sparser
"Our previous research has demonstrated that adult T-cell leukemia/lymphoma (ATLL) had high level of CYLD phosphorylation, down-regulating CYLD phosphorylation could increase CYLD deubiquitinase activity and then promoted apoptosis in ATLL [ xref ]."
33827598
sparser
"Therefore, we hypothesized that BTKis could promote the apoptosis of non-GCB-DLBCL by regulating the phosphorylation of CYLD, especially in rituximab resistant cases, and we proved this hypothesis through both in vivo and in vitro experiments."
33827598
sparser
"Previous work demonstrated that TNF-α induces IKKα− and IKKβ−mediated serine phosphorylation of CYLD (Ser 418) ."
20347313
sparser
"These results imply that the signal-stimulated CYLD phosphorylation negatively regulates its DUB function, thereby allowing temporary accumulation of ubiquitin chains on its targets, such as TRAF2."
19373246
sparser
"In vitro experiments using purified proteins demonstrated direct phosphorylation and activation of CYLD by the beta catalytic subunit of IKK."
24928390
sparser
"This reduces CYLD phosphorylation, activating this enzyme to remove more ubiquitin molecules from RIPK1, thus setting up a positive-feedback loop to deubiquitinate RIPK1 to the level allowing its association with the DISC."
32024820
rlimsp
"Phosphorylated peptides of CYLD detected under CaMKII activating conditions."
24614225
sparser
"These results suggested that BTKis inhibited tumor development in non-GCB-DLBCL mice model by down-regulating CYLD phosphorylation,which further confirmed that CYLD phosphorylation is an essential mediator in BTKis downstream pathway to promote tumor cell death in non-GCB-DLBCL in vivo."
33827598
sparser
"Overexpression of EGFR and ERBB2 can sequester β-catenin in EAC, and catenin phosphorylation by tyrosine kinase receptors prevents E-cadherin from binding."
15297940
sparser
"In addition to down-regulation of the NF-κB pathway [ xref ], CYLD phosphorylation has been demonstrated to have a physiological relevance in increasing cell transformation [ xref ], hence precise identification of a kinase or a kinase cascade involved in this process might provide potential targets for pharmacological intervention strategies in the treatment of cancer."
21480003
sparser
"We investigated CYLD phosphorylation by Western blotting with an antibody specific to CYLD phospho-Ser-418 ( xref )."
22424773
sparser
"Interestingly, pre-incubation of hippocampal cultures with IKK16 neither reduced NMDA-induced CYLD phosphorylation ( xref ) nor prevented recruitment of more CYLD to the PSD ( xref )."
24928390
sparser
"SMAC mimetics can similarly disrupt CYLD phosphorylation and lead to ATLL cell death through reduction of RIPK1 ubiquitination, which is CYLD dependent."
32024820
sparser
"Phosphorylation of the tumor suppressor CYLD by the breast cancer oncogene IKKepsilon promotes cell transformation."
19481526
sparser
"Other known substrates of IKKɛ are Akt, a mediator of cell survival, xref , xref , xref and the transcription factor cJun. xref Recently, IKKɛ was identified as an oncogene in breast cancer, xref where it was shown to phosphorylate the tumor suppressor CYLD, xref leading to cellular transformation."
25133483
sparser
"Interestingly, recent findings also suggest that OTULIN deficiency results in spontaneous inhibitory phosphorylation of CYLD, thereby limiting CYLD activity, which can be rescued by LUBAC inhibition [ xref ]."
33288901
sparser
"Whether CYLD phosphorylation is also a therapeutic target for non-GCB-DLBCL is unknown."
33827598
sparser
"These data demonstrate that phosphorylated CYLD exhibits less deubiquitinating activity than unphosphorylated CYLD."
19481526
sparser
"The phosphorylation of CYLD may be considered a mechanism of CYLD/IKK mutual regulation."
19373246
sparser
"BTK inhibitor ibrutinib down-regulated CYLD phosphorylation and inhibited tumor growth in xeograft mice model."
33827598
sparser
"Activation of NF-κB therefore appears to be one important mechanism by which CYLD phosphorylation regulates cell transformation."
19481526
sparser
"We have previously reported CaMKII-mediated phosphorylation of CYLD in isolated PSDs in the presence of Ca 2+ , with concomitant increase of DUB activity [ xref ]."
24928390
sparser
"In this study, we demonstrated that CYLD phosphorylation is a potential regulator for non-GCB-DLBCL development in vitro and in vivo."
33827598
sparser
"A CaMKII inhibitor CN21 had no effect on CYLD phosphorylation in the absence of Ca 2+ , but two different IKK inhibitors, IKK16 and tatNEMO, inhibited its phosphorylation."
24928390
sparser
"In the present study, we set out to identify the protein kinase responsible for the phosphorylation of CYLD in the absence of Ca 2+ ."
24928390
sparser
"The ubiquitination of CYLD can be regulated by phosphorylation, suggesting that the regulation of CYLD phosphorylation can be a potential mechanism to promote the apoptosis of hematological tumors."
33827598
sparser
"Indeed, the presence of NEMO is critical for the phosphorylation of CYLD by IKK catalytic subunits."
19373246
sparser
"However, when CYLD phosphorylation is suppressed by IKK inhibitors, its enzymatic activity is then turned on to remove ubiquitin from RIPK1, allowing its association with death-signaling molecules."
32024820
rlimsp
"Interestingly, in a large panel of HTLV1-transformed T-cell lines, CYLD is constitutively phosphorylated."
21824392
sparser
"To verify the effect of BTKis on the phosphorylation of CYLD and the apoptosis in vivo and in rituximab resistant non-GCB-DLBCL, the xeograft model and rituximab resistant non-GCB-DLBCL cells were generated by tumor cell inoculation and escalation of drug concentrations, respectively."
33827598
rlimsp
"Whether IKKε is involved in the CYLD phosphorylation in HTLV1-trnasformed T cells is yet to be investigated."
21824392
sparser
"Interestingly, in a large panel of HTLV1-transformed T-cell lines, CYLD is constitutively phosphorylated."
21824392
sparser
"However, the CYLD phosphorylation band in total cell lysates is masked by the phosphorylation of another protein of the same size which is also detected by the phospho(Ser418)-CYLD-specific antibody."
29755980
sparser
"A similar result was observed in vivo when we evaluated the contribution of CYLD phosphorylation to IKKε-induced tumorigenicity."
19481526
rlimsp
"The control peptide had no appreciable effect on the Ca2+/calmodulin-dependent phosphorylation of CYLD (Fig. 2B, lane 5)."
24614225
sparser
"However, there was no difference in CYLD phosphorylation between wild-type and IKKε knockout cells."
29755980
rlimsp
"CYLD activity is regulated by cleavage and phosphorylation."
26085216
sparser
"Incubating purified CYLD and purified CaMKII promotes phosphorylation on the same serine residues, providing evidence that the phosphorylation of CYLD in the PSD fractions is directly mediated by the endogenous CaMKII."
24614225
sparser
"Thus, IKKε regulates the deubiquitination of TRAF2 by phosphorylating CYLD."
19481526
reach
"Our finding appears to be in contradiction with previous reports, indicating that IKK mediated phosphorylation of CYLD promotes ubiquitination of TRAF2 in HeLa and HEK cells [XREF_BIBR], [XREF_BIBR] and of IKKgamma in HEK cells [XREF_BIBR]."
24928390
sparser
"Since knocking down all the kinases involved with hairpins is technically cumbersome, we undertook a dominant-negative approach by stably transfecting MT4 cells with a kinase-inactive TBK1-K38A mutant with the rationale that this would block all endogenous kinases from phosphorylating CYLD."
32024820
sparser
"In both samples, CYLD phosphorylation was elevated concomitant with that of TBK1/IKKε and IKKα/β (Supplementary Fig. xref )."
32024820
sparser
"Increased CYLD phosphorylation is a frequent event in ATLL."
32024820
sparser
"Interestingly, the CYLD phosphorylation requires IkappaB kinase gamma (IKKgamma) and can be induced by IKK catalytic subunits."
15870263
sparser
"In addition, two more Tax positive cell lines (MT2 and SLB1) showed increased levels of CYLD phosphorylation (Fig. xref )."
32024820
sparser
"In another case, inhibitory phosphorylation of CYLD after TNF-α stimulation is required for the accumulation of one of its proposed substrates, K63-ubiquitinated TRAF2."
16325574
rlimsp
"Here we provide evidence that inducible phosphorylation of CYLD is an important mechanism of its regulation."
15870263
sparser
"As MRT67307 blocks both IKKε and TBK1, we wanted to determine which of the two kinases is responsible for CYLD phosphorylation."
29755980
rlimsp
"As previously observed in mitogen-stimulated T cells, the upper band represented the phosphorylated CYLD, since it was converted to the basal form upon in vitro phosphatase treatment (Figure 4)."
21824392
sparser
"Thus, both the inhibitor and dominant-negative studies showed a correlation between reduction of CYLD phosphorylation and reduced cellular viability of MT4 cells."
32024820
rlimsp
"It also remains to be examined whether CYLD phosphorylation also contributes to HTLV1-induced T-cell transformation."
21824392
sparser
"CYLD becomes phosphorylated as a response to treatment with a number of NF-κB-inducing factors, such as LPS or TNF-α [ xref ]."
21480003
rlimsp
"Incubating purified CYLD and purified CaMKII promotes phosphorylation on the same serine residues, providing evidence that the phosphorylation of CYLD in the PSD fractions is directly mediated by the endogenous CaMKII."
24614225
rlimsp
"Interestingly, the CYLD phosphorylation requires IkappaB kinase gamma (IKKgamma) and can be induced by IKK catalytic subunits."
15870263
sparser
"We investigated CYLD phosphorylation by western blotting with an antibody specific to CYLD phospho-Ser-418 ( Figure 4 A )."
22424773
sparser
"Interestingly, we could detect a band with the phospho(Ser418)-CYLD-specific antibody in the total lysate control of the IKKε/TBK1 knock-out clones (Figures xref , xref ), indicating that this antibody specifically detects phosphorylated CYLD under native conditions, while it additionally detects an unknown protein of the same size in denaturing conditions."
29755980
sparser
"Here we could show that 5 μM MRT67307 completely prevented CYLD phosphorylation, while 1 or 2 μM reduced the phospho(Ser418)-CYLD signal only partially (Figure xref )."
29755980
sparser
"Conversely, disrupting CYLD phosphorylation using IKK inhibitors reactivates CYLD, which in turn, deubiquitinates and switches RIPK1 into a death-signaling molecule (Fig. xref )."
32024820
sparser
"IκB kinase (IKK), with regulatory subunit IKKγ, phosphorylates CYLD and results in the loss of its deubiquitinating function towards TRAF2( xref )."
23797609
sparser
"But whether BTKis can regulate CYLD phosphorylation and to improve cell apoptosis in hematopoietic malignancies, especially in non-GCB-DLBCL, is also unknown."
33827598
sparser
"Activation of IKK in isolated PSDs also promoted phosphorylation of CYLD and an increase in endogenous deubiquitinase activity specific for K63-linked polyubiquitins."
24928390
sparser
"Finally, our results also reveal that TCR stimulation results in the IKKε/TBK1-independent phosphorylation of an unknown 110 kDa protein, which can be inhibited by MRT67307, and which masks the detection of phosphorylated CYLD upon western blotting with a phospho(Ser418)-CYLD-specific antibody."
29755980
sparser
"But there were significant difference in phospho-BTK and phospho-CYLD in the non-GCB-DLBCL patient samples (P1–P5) compared with the lymphadenitis patient samples (B1–B5) (Fig.  xref a)."
33827598
sparser
"Proteins were extracted from the tumor tissue and Western blotting showed that the expression level of CYLD phosphorylation was significantly lower in BTKis group compared with in control group (Fig.  xref c)."
33827598
rlimsp
"Activation of endogenous CaMKII in isolated PSDs promotes phosphorylation of CYLD."
24614225
sparser
"These results indicated that phosphorylated CYLD is a crucial regulator of ATLL survival and a potential novel therapeutic target for pharmacologic modification in ATLL ( xref )."
34177595
sparser
"Disruption in RIPK1 ubiquitination is also known to disrupt IKK activity xref , xref , and therefore we hypothesized that SMAC mimetics could also reduce IKK activity and CYLD phosphorylation."
32024820
sparser
"While it has been reported that IKKα and IKKβ can phosphorylate Ser418 of CYLD ( xref ), we demonstrated that CYLD is phosphorylated much more efficiently by IKKε than by either of the canonical IKKs."
19481526
sparser
"Here we provide evidence that inducible phosphorylation of CYLD is an important mechanism of its regulation."
15870263
sparser
"In the present study we show that co-expression of CYLD and IKKε/TBK1 in HEK293T cells, as well as incubation with recombinant IKKε/TBK1, results in CYLD phosphorylation that can be detected with a phospho(Ser418)-CYLD-specific antibody upon immunoblotting."
29755980
sparser
"In all our experiments, we used lysates from Jurkat T cells (clone 3T8) xref as the negative control because of this cell line’s low basal levels of CYLD phosphorylation."
32024820
sparser
"MT4 cells stably transfected with TBK1-K38A had reduced CYLD phosphorylation (Fig. xref ) and a significantly slower growth rate (Fig. xref ) compared to cells transfected with a control gene."
32024820
sparser
"Unexpectedly, a phospho(Ser418)-CYLD signal was still detectable upon PMA/I stimulation in the IKKε/TBK1 double deficient Jurkat cells, indicating that a kinase different from IKKε and TBK1 is responsible for CYLD phosphorylation (Figure xref )."
29755980
rlimsp
"Since a phospho-mimetic CYLD mutant is defective in inhibiting Tax ubiquitination, the CYLD phosphorylation may contribute to the chronic activation of IKK and NF-κB in HTLV1-transformed T cells."
21824392
sparser
"The reported mechanisms underlying the alterations in gene expression include; (1) direct recruitment of a transcriptional repressor to the promoter, (2) serine phosphorylation of CYLD by IKK-ɛ, whi[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
24075517
sparser
"The observation that CYLD can also be phosphorylated in the absence of Ca 2+ in isolated PSDs led us to further explore the regulation of CYLD under basal conditions."
24928390
rlimsp
"Furthermore, Tax and CYLD appear to mutually regulate, since CYLD is constitutively phosphorylated in HTLV1-transformed T cells."
21824392
sparser
"We propose that phosphorylated CYLD, together with SPATA2 and LUBAC, functions as a ubiquitin-editing complex that balances Lys63- and Met1-linked polyubiquitin at receptor signaling complexes to promote LUBAC signaling."
34610306
sparser
"Thus, IKK-mediated CYLD phosphorylation deconjugates ubiquitin chains on RIPK1, increasing its kinase activity for the death signaling pathway."
35109788
sparser
"CYLD phosphorylation and cleavage caspase3 were detected by Western blotting, and the results further indicated that BTK inhibitors could enhance the apoptosis of non-GCB-DLBCL cells induced by rituximab through down-regulating the phosphorylation of CYLD (Fig.  xref g)."
33827598
sparser
"In the light of these data, it would be more rational to develop a new model between IKK-mediated CYLD phosphorylation and NRF2 activity."
35109788
rlimsp
"To examine the effect of phosphatase(s) on CYLD phosphorylation, PSD fractions (54 µg protein in a final volume of 90 µL) were incubated under phosphorylation conditions as described above, except that Microcystin-LR was excluded in one sample as indicated."
24614225
sparser
"An IKK inhibitor reduced lipid deposition, ROS production, CYLD phosphorylation levels and ΔΨm in vitro, which were reversed by knockdown of CYLD."
34711178
sparser
"These results demonstrate that CYLD phosphorylation is elevated in human ATLL."
32024820
sparser
"Phosphorylation of the tumor suppressor CYLD by the breast cancer oncogene IKKε promoted cell transformation ( xref )."
21170083
sparser
"Thus, CYLD undergoes constitutive phosphorylation in HTLV1-transformed T cells."
21824392
sparser
"In response to cellular stimuli, CYLD undergoes rapid and transient phosphorylation, which is required for signal-induced TRAF2 ubiquitination and activation of downstream signaling events."
15870263
sparser
"MRT67307 significantly decreased CYLD phosphorylation in wild-type cells (Figure xref )."
29755980
sparser
"To determine whether phosphorylation of CYLD affects RIP2-induced NEMO ubiquitination, HEK-293T cells were cotransfected with Myc-NEMO, OMNI-RIP2, HA-Ubiquitin, GFP-CYLD, and GST-IKKε."
19481526
sparser
"Our results indicated that CYLD phosphorylation were significant different between non-GCB-DLBCL samples and control samples."
33827598
sparser
"Additionally, phosphorylation of CYLD was found to downregulate CYLD activity: transient phosphorylation by IKK in a serine cluster just upstream of the TRAF2 binding site, attenuates DUB function xref ."
23335970
sparser
"We first analyzed CYLD phosphorylation in C8166 and MT4 T cell lines, which are HTLV-1-transformed T cells."
32024820
sparser
"Our previous study indicated high protein expression levels of CYLD phosphorylation in adult T-cell leukemia/lymphoma (ATLL) and regulating CYLD phosphorylation could regulate CYLD ubiquitin activity to improve ATLL cells apoptosis [ xref ]."
33827598
sparser
"Biochemical and genetic evidence suggests that IKK is a kinase that mediates CYLD phosphorylation. xref As CYLD physically interacts with the IKK regulatory subunit, NEMO, it is likely that CYLD is recruited to the IKK holoenzyme by NEMO."
19373246
rlimsp
"CaMKII co-immunoprecipitates with CYLD and promotes phosphorylation of CYLD at the PSD."
24614225
sparser
"In primary ATLL samples and in cell line models, an increased baseline level of CYLD phosphorylation was observed."
32024820
sparser
"To examine regulation of CYLD phosphorylation in intact cells, immunoEM studies were carried out in hippocampal cultures using an antibody specific for p-CYLD."
24928390
sparser
"As shown in xref , knocking down CDK6 in EAC cells reduced phosphorylation of pRB at serines 780 and 795 (both are known targets of CDK6/Cyclin D1 complex ( xref )) although with much lesser effect in OE33."
21593195
sparser
"The research on how CYLD phosphorylation regulate the occurrence and development of hematological malignances is still lacking."
33827598
sparser
"Phosphorylation of CYLD under basal conditions was inhibited by IKK16."
24928390
sparser
"We have previously shown that induction of CYLD phosphorylation by TNF-α or mitogens is mediated by the IKK complex [ xref ]."
21824392
sparser
"Interestingly, IKKε had previously been reported to phosphorylate and inactivate CYLD, raising the possibility that the negative regulation of CYLD by IKKε also has a role in the prevention of RIPK1 dependent cell death in the context of inflammation as well ( xref )."
31316982
rlimsp
"Activation of IKK in isolated PSDs also promoted phosphorylation of CYLD and an increase in endogenous deubiquitinase activity for K63-linked polyubiquitins."
24928390
sparser
"Western blotting showed that in OCI-Ly10S cells, BTK inhibitor PCI-32765 could enhance the apoptosis of non-GCB-DLBCL cells induced by rituximab through down-regulating the phosphorylation of CYLD (Fig.  xref e)."
33827598
sparser
"Furthermore, CYLD phosphorylation is required for TRAF2 ubiquitination and downstream activation of NF-κB and JNK [ xref ]."
21119682