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USP14 is phosphorylated. 79 / 79
| 53 26
sparser
"In addition, treatment with lipopolysaccharide (LPS) induced the serine phosphorylation of USP14, further deregulating the levels of I-κB in USP14-overexpressing cells ( xref ) ( xref )."
35069211
sparser
"Interestingly, we found that USP14 was associated with RelA, a binding partner of I-κB, suggesting that RelA is the linker between USP14 and I-κB. Lipopolysaccharide (LPS) treatment induced serine phosphorylation of USP14 as well as further reducing I-κB levels in HA-USP14-overexpressed MLE12 cells as compared with empty vector transfected cells."
23615914
rlimsp
"We also demonstrate that phosphorylation of USP14 is critical for Akt to control UPS and consequentially global protein degradation via the UPS."
26523394
sparser
"1) We analyzed the distribution of phosphorylated and total USP14 by glycerol gradient centrifugation (Koulich et al., 2008) and found that phosphorylated USP14 was largely present in fractions without proteasome while unphosphorylated USP14 was found in the same fractions with proteasome ( xref )."
26523394
rlimsp
"The reaction products were resolved by SDS-PAGE, and USP14 phosphorylation was detected using an antibody that specifically recognizes Ser432 phosphorylation of USP14 or determined by differential migration on phos-tag gels."
26523394
sparser
"Is it only the proteasome bound fraction of USP14 that is phosphorylated or is it the unbound USP14 that is phosphorylated."
26523394
rlimsp
"To further characterize the effect of Ser432 phosphorylation, we expressed and purified recombinant S432E USP14 protein, which mimics the phosphorylation state of USP14, from E. coli (Figure 3—figure supplement 1) and analyzed its activity by Ub-AMC assay."
26523394
sparser
"The phos-tag gels suggest that only a minor fraction of USP14 is phosphorylated."
26523394
sparser
"These observations may suggest that phosphorylated Usp14 could “moonlight” as an active enzyme outside of the proteasome."
28988953
rlimsp
"Regulation of UPS by Akt depends on phosphorylation of USP14."
26523394
sparser
"Phosphorylation of USP14 regulates global protein degradation."
26523394
sparser
"2) xref reports that the endogenous USP14 is phosphorylated in PTEN KO cells in the absence of IGF1 stimulation."
26523394
sparser
"Finally they report that the phosphorylation of USP14 suppresses the activity of the proteasome in cells."
26523394
rlimsp
"(I) Akt regulates UPS function through phosphorylation of USP14."
26523394
sparser
"Since Akt is dramatically activated in PTEN-deficient cancer cells, the control of USP14 phosphorylation by Akt may provide a mechanism for cancer cells with PTEN loss, one of the most common cancer mutations, to control global intracellular proteostasis by regulating protein degradation through proteasomes."
26523394
rlimsp
"Phosphorylation stimulates ubiquitin-specific protease-14 (USP14) activity towards both K48 and K63 ubiquitination."
26523394
rlimsp
"Conversely, overexpression of USP14 increased the expression of AR and PSA and decreased the expression of MDM2 and its phosphorylation (Figure 5e)."
28151478
rlimsp
"d NiPT inhibits the phosphorylation of USP14."
27884201
sparser
"A previous study has reported that phosphorylation and activation of USP14 was regulated by AKT."
31013812
rlimsp
"Phosphorylation of USP14 regulates global protein degradation."
26523394
sparser
"As requested, this experiment was performed to show that phosphorylation of USP14 in PTEN KO cells was indeed suppressed by MK2206 and AZD5363 ( xref )."
26523394
sparser
"Furthermore, the Akt-mediated phosphorylation and activation of USP14 may provide a mechanism for promoting tumorigenesis in PTEN negative cancer cells ( Xu et al., 2015 )."
27592452
rlimsp
"Akt regulates ubiquitin–proteasome system (UPS) function through phosphorylation of ubiquitin-specific protease-14 (USP14)."
26523394
rlimsp
"Phosphorylation of ubiquitin-specific protease-14 (USP14) regulates global protein degradation."
26523394
sparser
"We also demonstrate that phosphorylation of USP14 is critical for Akt to control UPS and consequentially global protein degradation via the UPS."
26523394
sparser
"Our study reveals a novel mechanistic connection between activated Akt and the UPS through regulating of USP14 phosphorylation, which can impact global proteostasis in PTEN-negative cancer cells."
26523394
sparser
"Since Akt can be activated by a wide range of growth factors and is under negative control by phosphoinosotide phosphatase PTEN, we suggest that regulation of UPS by Akt-mediated phosphorylation of USP14 may provide a common mechanism for growth factors to control global proteostasis and for promoting tumorigenesis in PTEN-negative cancer cells."
26523394
sparser
"Phosphorylated Ser432 Usp14 was detected in proteasome-free fractions using glycerol gradient centrifugation, whereas unphosphorylated Usp14 was mostly found in the proteasome fractions [ xref ]."
28988953
sparser
"Taken together, our results suggest that phosphorylation of USP14 may provide a mechanism for Akt to regulate global protein degradation through the proteasome, which in turn may control key cellular pathways involved in regulating metabolism and tumorigenesis."
26523394
sparser
"Both mechanisms involve AKT1, which exhibits yet another way of influencing BECN1 by phosphorylating USP14, which removes K63-linked ubiquitins from BECN157."
31852899
sparser
"They are therefore candidates to phosphorylate USP14, but their activation is not blocked by MK2206."
26523394
rlimsp
"Notably, the phosphorylation levels of USP14 were decreased in cells when treated with MK2206, an inhibitor of Akt (Figure 2D), or when serum deprived, a condition known to inactivate endogenous Akt (Zhang et al., 2015) (Figure 2D)."
26523394
sparser
"Regulation of UPS by Akt depends on phosphorylation of USP14."
26523394
rlimsp
"Specifically, Usp14 regulates the ubiquitination of Dvl and its subsequent phosphorylation, which is essential for activation of downstream Wnt signaling."
23958854
sparser
"Interestingly, Akt was shown to phosphorylate USP14, enhancing its deubiquitinating activity on K48 and K63 ubiquitin linkages, thereby promoting tumor development and cisplatin resistance as well as DSB repair dynamics over regulation of RNF168 mediated ubiquitination [ xref , xref ]."
33202866
sparser
"Furthermore, the phosphorylation of USP14 (S241) was substantially reduced in NiPT-treated CML cells, also indicative of suppression of the DUB activity of USP14 by NiPT treatment (Fig.  xref ) [ xref ]."
27884201
rlimsp
"Active-site-directed labeling of proteasomal DUBs, as well as the phosphorylation of USP14 was used for evaluating the inhibition of the DUBs activity by NiPT."
27884201
sparser
"Our results do not exclude the possibility that other kinases that are activatable by growth factors and activated in PTEN-deficient conditions can also mediate the phosphorylation of USP14."
26523394
sparser
"Emerging evidence also suggest that the BDNF signaling pathway may regulate not only the activity of the 26S proteasome but also the phosphorylation of USP14 ()."
29407217
sparser
"USP14 can be phosphorylated and activated by the protein kinase Akt, thus negatively regulating autophagy by removing K63-linked ubiquitination from Beclin 1 [ xref ]."
31883910
rlimsp
"Taken together, our results suggest that phosphorylation of USP14 may provide a mechanism for Akt to regulate global protein degradation through the proteasome, which in turn may control key cellular pathways involved in regulating metabolism and tumorigenesis."
26523394
sparser
"Since UPS is critically involved in the degradation of cellular short-lived proteins which frequently serve in mediating intracellular signaling process, the regulation of UPS by Akt-mediated phosphorylation of USP14 may provide a mechanism to control multiple signaling process, raising the possibility that control of short-lived proteins might serve as an active process that has impact on cellular signaling, rather than as a degradative process per se."
26523394
rlimsp
"Phosphorylation of USP14 promotes both K48 and K63 deubiquitination activity."
26523394
sparser
"Both mechanisms involve AKT1, which exhibits yet another way of influencing BECN1 by phosphorylating USP14, which removes K63-linked ubiquitins from BECN1 xref ."
31852899
sparser
"We also demonstrate that phosphorylation of USP14 is critical for Akt to regulate proteasome activity and consequently global protein degradation."
26523394
sparser
"Similar to the inhibition of USP14, blocking AKT, which mediates the phosphorylation of USP14, was also reported to rescue the activity of NHEJ-DDR proteins during autophagy in PTEN-deficient cells ( xref )."
35069211
rlimsp
"To further understand the physiological roles of Akt-mediated USP14 phosphorylation and subsequently activation, we sought to study the impact of USP14 phosphorylation on global protein degradation."
26523394
rlimsp
"The enrichment of phosphorylated peptides by TiO2 was performed with tryptic USP14 peptides. The enriched phosphorylated peptides were analyzed on Orbitrap Fusion mass spectrometer (Thermo Scientific)."
26523394
sparser
"Moreover, no selective inhibitors towards phosphorylated USP14 have been identified to date."
35069211
sparser
"To further understand the physiological roles of Akt-mediated USP14 phosphorylation and subsequently activation, we sought to study the impact of USP14 phosphorylation on global protein degradation."
26523394
sparser
"In addition, former study stated that activating AKT induced phosphorylation and activation of USP14. xref Since we have proved that SPAG5‐AS1 positively regulated AKT/mTOR pathway in HG‐treated HPCs, we tried to test whether SPAG5‐AS1 could induce USP14 phosphorylation."
31957155
sparser
"It was demonstrated that AKT-mediated phosphorylation of USP14 may provide a novel mechanism to control multiple signaling processes, including inhibition of the UPS through USP14 activation [ xref ]."
| PMC
rlimsp
"Our results do not exclude the possibility that other kinases that are activatable by growth factors and activated in PTEN-deficient conditions can also mediate the phosphorylation of USP14."
26523394
sparser
"Active-site-directed labeling of proteasomal DUBs, as well as the phosphorylation of USP14 was used for evaluating the inhibition of the DUBs activity by NiPT."
27884201
rlimsp
"Since UPS is critically involved in the degradation of cellular short-lived proteins which frequently serve in mediating intracellular signaling process, the regulation of UPS by Akt-mediated phosphorylation of USP14 may provide a mechanism to control multiple signaling process, raising the possibility that control of short-lived proteins might serve as an active process that has impact on cellular signaling, rather than as a degradative process per se."
26523394
sparser
"Indeed, phosphorylation of Usp14 (or the use of a Usp14 phosphomimetic) results in increased activity in assays with the fluorogenic substrate ubiquitin-7-amino-4-methylcoumarin (Ub-AMC), for both Usp[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
28988953
rlimsp
"Our study reveals a novel mechanistic connection between activated Akt and the UPS through regulating of USP14 phosphorylation, which can impact global proteostasis in PTEN-negative cancer cells."
26523394
sparser
"Phosphorylation of USP14 promotes both K48 and K63 deubiquitination activity."
26523394
sparser
"To address this question from the reviewers, we performed glycerol density gradient centrifugation (Koulich et al., 2008) to determine the distribution of free and proteasome-associated phosphorylated USP14 inPten -/- MEFs (with high USP14 S432 phosphorylation) cells lysates ( xref )."
26523394
sparser
"Previously, we showed that LPS stimulates the serine phosphorylation of USP14 ( xref )."
28611184
sparser
"To explore the role of PTEN in AKT-mediated USP14 phosphorylation in NHEJ repair, we examined IRIF by USP14 in VCaP (PTEN+) versus LnCaP (PTEN-) cells following IR."
31740976
rlimsp
"We also demonstrate that phosphorylation of USP14 is critical for Akt to regulate proteasome activity and consequently global protein degradation."
26523394
sparser
"Indeed, phosphorylation of Usp14 (or the use of a Usp14 phosphomimetic) results in increased activity in assays with the fluorogenic substrate ubiquitin-7-amino-4-methylcoumarin (Ub-AMC), for both Usp14 in isolation and Usp14 in the proteasome, suggesting that phosphorylation and proteasome binding activate Usp14 via unrelated mechanisms [ xref ]."
28988953
sparser
"We next tested the importance of USP14 phosphorylation for Akt to regulate UPS."
26523394
sparser
"We identify AKT-mediated phosphorylation of USP14 as a negative regulator of NHEJ-DDR in autophagy-deficient cells."
31740976
sparser
"By regulating the rate of proteasomal degradation through phosphorylating USP14, Akt may exert controls over the entire proteome of short-lived proteins as well as cellular protein quality."
26523394
sparser
"In addition, Ser432 is surrounded by a cluster of highly negatively charged residues, which electrostatically repulse the negatively charged phosphate group of the phosphorylated USP14, thus leading to the rearrangement of the BL2 loop, disturbing the inhibitory effect of BL2 on USP14 ( xref ) ( xref )."
35069211
sparser
"A previous study has reported that serine phosphorylation of USP14 showed high DUB activity ( xref )."
32391376
rlimsp
"To assess the impact of USP14 phosphorylation on its selectivity towards different types of ubiquitin linkages, we incubated USP14 WT and S432E mutant protein with diubiquitin species of K48, K63, and linear linkages."
26523394
sparser
"Furthermore, the phosphorylation of USP14 (S241) was substantially reduced in PtPT-treated CML cells ( xref ), also indicative of suppression of the DUB activity of USP14 by PtPT treatment. xref "
28682311
sparser
"AKT, which mediates Ser432-USP14 phosphorylation, was required for IRIF formation by USP14."
31740976
sparser
"To determine if Ser143 and Ser432 were indeed phosphorylated by Akt, we used this pan phospho-Ser antibody as above and found phosphorylation of wild type (WT) USP14, but not of S143A/S432A mutant USP14, after incubating with activated Akt in a kinase assay ( xref )."
26523394
sparser
"Accumulation of ubiquitinated proteins, as well as the decrease of phosphorylated USP14 were observed as early as 6 h during the course of treatment ( xref )."
28682311
sparser
"Phosphorylated Usp14 was detected in proteasome-free fractions using glycerol gradient centrifugation, whereas unphosphorylated Usp14 was mostly found in the proteasome-containing fractions [74] ."
28988953
rlimsp
"By regulating the rate of proteasomal degradation through phosphorylating USP14, Akt may exert controls over the entire proteome of short-lived proteins as well as cellular protein quality."
26523394
rlimsp
"In this study, we report that USP14 is an Akt substrate and that this phosphorylation activates the DUB activity of USP14 both in vitro and in cells."
26523394
sparser
"Moreover, the DSB repair function of USP14 is dependent on Akt-mediated phosphorylation and activation of USP14."
31740976
rlimsp
"The differential migration of phospho-USP14 on phos-tag-containing gels was determined as shown in the bottom panel."
26523394
sparser
"In addition, LPS treatment induced serine phosphorylation of USP14 quickly ( xref )."
32391376