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VCPIP1 is phosphorylated. 76 / 76
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"In addition to its S130 phosphorylation, VCIP135 is also phosphorylated at multiple sites at the C-terminus, which regulates VCIP135 membrane association and p97 interaction ( xref )."
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"These results suggest that VCPIP1 is phosphorylated by ATM/ATR following DPC induction."
32649882
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"Our in vitro Golgi reassembly assay confirmed that VCIP135 phosphorylation at S130 attenuates p97/p47-mediated Golgi membrane fusion ( xref )."
25904330
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"Given our results, we propose that VCIP135 phosphorylation on S130 in early mitosis inhibits its activity and blocks Golgi membrane fusion, resulting in mitotic Golgi fragmentation; while VCIP135 reactivation by S130 dephosphorylation in telophase initiates p97/p47-mediated Golgi membrane fusion for postmitotic Golgi reassembly."
25904330
rlimsp
"VCIP135 is phosphorylated at multiple sites during mitosis (Totsukawa ; Zhang ), but it was not clear whether and how VCIP135 phosphorylation regulates its DUB activity and function in Golgi membrane fusion."
25904330
sparser
"As presented in Fig. 2 A, Cdc2-mediated VCIP135 phosphorylation inhibited its binding to GST-p97 (upper panel, lane 2)."
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rlimsp
"Mitotic phosphorylation of VCIP135 blocks p97ATPase-mediated Golgi membrane fusion."
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sparser
"These results demonstrate that VCIP135 is phosphorylated in mitosis on S130 and is dephosphorylated in later mitosis."
25904330
sparser
"Taken together, our results demonstrate that phosphorylation of VCIP135 on S130 in early mitosis inactivates its DUB activity for mitotic Golgi disassembly; dephosphorylation in telophase reactivates VCIP135 to promote p97/p47-mediated postmitotic Golgi membrane fusion."
25904330
sparser
"Mitotic phosphorylation of VCIP135 at S130 inhibits its deubiquitinase activity."
25904330
rlimsp
"Therefore VCIP135 is phosphorylated in early mitosis and dephosphorylated in telophase."
25904330
sparser
"We next tested the binding of phosphorylated VCIP135 to WAC, as presented in Fig. 3 A. Cdc2-phosphorylated VCIP135 still bound to GST-WAC (upper panel, lane 2), suggesting that the phosphorylation of[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"On the other hand, neither PD98059 (MEK inhibitor), SB203580 (p38 MAP kinase inhibitor), KT5720 (protein kinase A inhibitor) nor calphostin C (protein kinase C inhibitor) had any effect on VCIP135 pho[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
23500464
sparser
"Therefore it is important to understand how VCIP135 DUB activity is accurately regulated to fuse mitotic Golgi membranes only at the end of mitosis and whether this regulation is through VCIP135 phosphorylation."
25904330
sparser
"The deubiquitinating enzyme activity of phosphorylated VCIP135 was also assayed."
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rlimsp
"Furthermore, VCIP135 is highly phosphorylated in mitosis."
25904330
sparser
"VCIP135 phosphorylation at multiple sites reduces its association with Golgi membranes and its interaction with p97, and this is thought to be a mechanism that restricts p97 function in the fusion of the postmitotic Golgi membrane ( xref )."
25904330
sparser
"We first tested whether phosphorylated VCIP135 binds to p97."
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rlimsp
"Although the C-terminus has no DUB activity (Figure 2A), phosphorylation of the C-terminus might also regulate VCIP135 DUB activity."
25904330
rlimsp
"Our results demonstrate that VCIP135 phosphorylation regulates its Golgi membrane association and p97 interaction, and thus contributes to the tight control of the Golgi disassembly and reassembly process during the cell cycle."
24163436
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"The phosphorylated VCIP135 was then used for binding experiments with GST-p97."
23500464
sparser
"VCPIP1 is phosphorylated by ATM/ATR upon DPC induction."
32649882
sparser
"Our results demonstrate that VCIP135 phosphorylation regulates its Golgi membrane association and p97 interaction, and thus contributes to the tight control of the Golgi disassembly and reassembly process during the cell cycle."
24163436
sparser
"Both the T760A and S767A mutation reduced VCIP135 phosphorylation by mitotic cytosol ( Fig. 1 E, lanes 3 and 4), while neither the S764A nor S997A mutation showed any effect (lanes 2 and 5)."
23500464
sparser
"In this study, we found that phosphorylation of VCIP135 at a single site, S130, is necessary and sufficient to inactivate its DUB activity for Golgi disassembly in early mitosis."
25904330
sparser
"For determination of the temporal control of VCIP135 phosphorylation during progression through the cell cycle, cells expressing Flag-tagged VCIP135 were synchronized in mitosis by nocodazole arrest (Mit) followed by release in fresh medium and were analyzed at different time points."
25904330
sparser
"Phosphorylation of VCIP135 at S130 attenuates p97/p47-mediated Golgi membrane fusion."
25904330
sparser
"VCIP135 is phosphorylated in mitosis ( xref ); therefore we hypothesized that the DUB activity of VCIP135 is regulated by phosphorylation."
25904330
sparser
"Taken together, these data suggest that VCPIP1 phosphorylation by ATM/ATR is important for VCPIP1 activation following DPC induction."
32649882
sparser
"We established that VCIP135 is mitotically phosphorylated at Threonine-760 and Serine-767 and that the phosphorylated VCIP135 cannot bind to p97."
23500464
sparser
"Therefore, Cdc2 was confirmed as the kinase responsible for VCIP135 phosphorylation."
23500464
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"When both the T760A and S767A mutations were induced into VCIP135 (VCIP135(T760A, S767A)), phosphorylation of VCIP135 by mitotic cytosol was inhibited (lane 6)."
23500464
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"To clarify the significance of this mitotic phosphorylation of VCIP135 in Golgi biogenesis, we performed the in vitro Golgi reassembly assay ( Fig. 4 ) [12] ."
23500464
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"These results demonstrate that phosphorylation of VCIP135 at S130 controls its deubiquitinase activit y in vivo."
25904330
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"Together with our finding that VCIP135 is phosphorylated at mitosis, the mitotic inhibition of the p97/p37-mediated and p97/p47-mediated Golgi assembly are thought to be achieved in multiple ways; by [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
23500464
sparser
"This observation led us to hypothesize that VCPIP1 phosphorylation affects its activity."
32649882
sparser
"Phosphomimetic mutants at known VCIP135 phosphorylation sites prevent the association of VCIP135 with p97 [ xref ]."
28819009
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"VCIP135 is phosphorylated on multiple sites at the C-terminus in addition to S130 ( xref ; xref ), and a total of 13 sites on VCIP135 are phosphorylated in mitosis (Supplemental Figure 1)."
25904330
sparser
"The phosphorylation of VCIP135(728–1221) was also suppressed by the Cdc2 inhibitor, olomoucine (right lane)."
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rlimsp
"For determination of the temporal control of VCIP135 phosphorylation during progression through the cell cycle, cells expressing Flag-tagged VCIP135 were synchronized in mitosis by nocodazole arrest (Mit) followed by release in fresh medium and were analyzed at different time points."
25904330
sparser
"We tested VCIP135(T760E, S767E), which mimics Threonine-760 and Serine-767 phosphorylation, instead of phosphorylated VCIP135."
23500464
rlimsp
"VCIP135 function requires membrane association and p97 interaction, both of which are inhibited in mitosis by VCIP135 phosphorylation."
24163436
sparser
"Determination of the phosphorylation of VCIP135 at mitosis led to the question as to what effect this phosphorylation has on the function of VCIP135."
23500464
sparser
"As presented in Fig. 1 B, VCIP135 was strongly phosphorylated in mitotic cytosol (lane 1)."
23500464
sparser
"VCIP135 function requires membrane association and p97 interaction, both of which are inhibited in mitosis by VCIP135 phosphorylation."
24163436
rlimsp
"Our previous study showed that phosphorylation at the C-terminus (e.g., 12E) inhibits VCIP135 Golgi membrane association and p97 interaction (Zhang )."
25904330
sparser
"Next we determined how VCIP135 phosphorylation at S130 regulates VCIP135's deubiquitinase activity during the cell cycle."
25904330
sparser
"Autoradiography revealed that VCIP135 was phosphorylated at mitosis ( Fig. 1 A, bottom panel)."
23500464
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"As shown in xref , following FA and CPT treatment, VCPIP1 was phosphorylated at its SQ/TQ motifs, which are consensus ATM/ATR phosphorylation sites."
32649882
sparser
"VCIP135 phosphorylated by Cdc2 generated almost the same amount of mono ubiquitin after the reaction as non-phosphorylated VCIP135 ( Fig. 3 C, top panel, lane 2)."
23500464
sparser
"VCIP135 was phosphorylated by incubation with Cdc2, and then its phosphorylated state was ‘frozen’ by addition of staurosporine and microcystin-LR."
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rlimsp
"These results suggest that p97/p47 membrane fusion activity is regulated by phosphorylation of VCIP135."
25904330
sparser
"Therefore VCIP135 is phosphorylated in early mitosis and dephosphorylated in telophase."
25904330
sparser
"Fig. 1 D shows that VCIP135(728–1221) was phosphorylated by mitotic cytosol (middle lane), while VCIP135(1–727) was not (left lane)."
23500464
rlimsp
"Therefore it is important to understand how VCIP135 DUB activity is accurately regulated to fuse mitotic Golgi membranes only at the end of mitosis and whether this regulation is through VCIP135 phosphorylation."
25904330
rlimsp
"Phosphorylation regulates VCIP135 function in Golgi membrane fusion during the cell cycle."
24163436
sparser
"Furthermore, VCIP135 is highly phosphorylated in mitosis."
25904330
sparser
"As Golgi disassembly and reassembly is temporally regulated during cell cycle progression, it is reasonable to speculate that VCIP135 phosphorylation and activity are also under precise control."
25904330
sparser
"To investigate whether there was mitotic phosphorylation of VCIP135, Hela cells were synchronized and incubated in medium containing 32 P-orthophosphate."
23500464
sparser
"We clarified that VCIP135, an essential factor in both p97 membrane fusion pathways, is phosphorylated on Threonine-760 and Serine-767 by Cdc2 at mitosis and that this phosphorylated VCIP135 does not bind to p97."
23500464
sparser
"Phosphorylation of Vcpip1 at pT760 by CDK1 inhibits Golgi membrane fusion, a process which is reported to be a prerequisite for entry into mitosis."
26643407
sparser
"This is also found with DUBs, as exemplified by the regulated expression and phosphorylation of A20, and the phosphorylation of VCIP135 ( xref ; xref ; xref )."
32107274
rlimsp
"As Golgi disassembly and reassembly is temporally regulated during cell cycle progression, it is reasonable to speculate that VCIP135 phosphorylation and activity are also under precise control."
25904330
sparser
"During early mitosis, phosphorylated VCIP135 is inactive and may not be able to bind p97."
28819009
rlimsp
"These results demonstrate that the function of VCIP135 in Golgi membrane fusion is tightly regulated by phosphorylation during the cell cycle, wherein phosphorylation at S130 and the C-terminus suppress DUB activity and membrane association, respectively, while dephosphorylation stimulates DUB activity, membrane association, and p97 interaction that are required for postmitotic Golgi reassembly."
25904330
sparser
"Staurosporine (broad serine/threonine kinase inhibitor) and olomoucine (Cdc2 inhibitor) inhibited VCIP135 phosphorylation (lanes 2 and 3)."
23500464
sparser
"The DUB activity was low when VCIP135 was highly phosphorylated at S130 in early mitosis ( xref , lanes 2–4); while the DUB activity increased significantly when S130 was dephosphorylated in late mitosis ( xref , lanes 5–7)."
25904330
sparser
"These results suggest that p97/p47 membrane fusion activity is regulated by phosphorylation of VCIP135."
25904330
sparser
"These results demonstrate that phosphorylation of VCIP135 at S130 attenuates p97/p47-mediated Golgi membrane fusion."
25904330
rlimsp
"Therefore the function of VCIP135 is tightly regulated by phosphorylation in which dephosphorylation of all the sites in both the N- and C-terminus is required to reactivate VCIP135 for p97/p47-mediated postmitotic Golgi membrane fusion."
25904330
sparser
"VCIP135 is phosphorylated at multiple sites during mitosis ( xref ; xref ), but it was not clear whether and how VCIP135 phosphorylation regulates its DUB activity and function in Golgi membrane fusion."
25904330
rlimsp
"VCIP135 phosphorylation at multiple sites reduces its association with Golgi membranes and its interaction with p97, and this is thought to be a mechanism that restricts p97 function in the fusion of the postmitotic Golgi membrane (Zhang )."
25904330
rlimsp
"VCIP135 is phosphorylated in mitosis (Zhang ); therefore we hypothesized that the DUB activity of VCIP135 is regulated by phosphorylation."
25904330
rlimsp
"Because VCIP135 is a deubiquitinase, regulation of its enzymatic activity is necessary to accurately control the timing of substrate cleavage. In the case of VCIP135, we found that it is highly phosphorylated in mitosis and identified S130 as the most critical site whose phosphorylation in early mitosis inactivates VCIP135 DUB activity."
25904330
sparser
"Nuclear VCPIP1 was phosphorylated at the SQ/TQ motif in wild-type but not ATM-deficient cells ( xref )."
32649882
sparser
"We next investigated the effect of several protein kinase inhibitors on the mitotic phosphorylation of VCIP135 ( Fig. 1 B, lanes 2–7)."
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