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VCPIP1 is phosphorylated. 59 / 78
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"We next investigated the effect of several protein kinase inhibitors on the mitotic phosphorylation of VCIP135 ( Fig. 1 B, lanes 2–7)."
23500464
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"Staurosporine (broad serine/threonine kinase inhibitor) and olomoucine (Cdc2 inhibitor) inhibited VCIP135 phosphorylation (lanes 2 and 3)."
23500464
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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
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"It is known that the phosphorylation of VCIP135 during mitosis disrupts its association with p97 ( xref , xref )."
38072049
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"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
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"VCPIP1 is phosphorylated by ATM/ATR upon DPC induction."
32649882
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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
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"Nuclear VCPIP1 was phosphorylated at the SQ/TQ motif in wild-type but not ATM-deficient cells ( xref )."
32649882
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"These results suggest that VCPIP1 is phosphorylated by ATM/ATR following DPC induction."
32649882
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"This observation led us to hypothesize that VCPIP1 phosphorylation affects its activity."
32649882
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"Taken together, these data suggest that VCPIP1 phosphorylation by ATM/ATR is important for VCPIP1 activation following DPC induction."
32649882
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"To investigate whether there was mitotic phosphorylation of VCIP135, Hela cells were synchronized and incubated in medium containing 32 P-orthophosphate."
23500464
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"Autoradiography revealed that VCIP135 was phosphorylated at mitosis ( Fig. 1 A, bottom panel)."
23500464
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"As presented in Fig. 1 B, VCIP135 was strongly phosphorylated in mitotic cytosol (lane 1)."
23500464
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"Therefore, Cdc2 was confirmed as the kinase responsible for VCIP135 phosphorylation."
23500464
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"Fig. 1 D shows that VCIP135(728–1221) was phosphorylated by mitotic cytosol (middle lane), while VCIP135(1–727) was not (left lane)."
23500464
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"The phosphorylation of VCIP135(728–1221) was also suppressed by the Cdc2 inhibitor, olomoucine (right lane)."
23500464
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"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
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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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"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
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"We previously showed that VCIP135 is phosphorylated on Thr-760 and Ser-767 by Cdc2 during mitosis and that the phosphorylation of VCIP135 decreases its binding affinity for p97 and inhibits p97-mediated Golgi membrane fusion during mitosis ( xref )."
38072049
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"We first tested whether phosphorylated VCIP135 binds to p97."
23500464
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"VCIP135 was phosphorylated by incubation with Cdc2, and then its phosphorylated state was ‘frozen’ by addition of staurosporine and microcystin-LR."
23500464
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"The phosphorylated VCIP135 was then used for binding experiments with GST-p97."
23500464
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"As presented in Fig. 2 A, Cdc2-mediated VCIP135 phosphorylation inhibited its binding to GST-p97 (upper panel, lane 2)."
23500464
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"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]"
23500464
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"Furthermore, VCIP135 is highly phosphorylated in mitosis."
25904330
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"The deubiquitinating enzyme activity of phosphorylated VCIP135 was also assayed."
23500464
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"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
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"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
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"These results suggest that p97/p47 membrane fusion activity is regulated by phosphorylation of VCIP135."
25904330
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"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
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"Mitotic phosphorylation of VCIP135 at S130 inhibits its deubiquitinase activity."
25904330
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"We established that VCIP135 is mitotically phosphorylated at Threonine-760 and Serine-767 and that the phosphorylated VCIP135 cannot bind to p97."
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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"We tested VCIP135(T760E, S767E), which mimics Threonine-760 and Serine-767 phosphorylation, instead of phosphorylated VCIP135."
23500464
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"VCIP135 is phosphorylated in mitosis ( xref ); therefore we hypothesized that the DUB activity of VCIP135 is regulated by phosphorylation."
25904330
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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
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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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"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
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"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
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"These results demonstrate that VCIP135 is phosphorylated in mitosis on S130 and is dephosphorylated in later mitosis."
25904330
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"Next we determined how VCIP135 phosphorylation at S130 regulates VCIP135's deubiquitinase activity during the cell cycle."
25904330
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"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
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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
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"Therefore VCIP135 is phosphorylated in early mitosis and dephosphorylated in telophase."
25904330
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"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
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"Phosphorylation of VCIP135 at S130 attenuates p97/p47-mediated Golgi membrane fusion."
25904330
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"These results demonstrate that phosphorylation of VCIP135 at S130 attenuates p97/p47-mediated Golgi membrane fusion."
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
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"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
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"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
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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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"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 )."
25904330
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"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
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"VCIP135 function requires membrane association and p97 interaction, both of which are inhibited in mitosis by VCIP135 phosphorylation."
24163436
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"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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"Phosphomimetic mutants at known VCIP135 phosphorylation sites prevent the association of VCIP135 with p97 [ xref ]."
28819009
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"During early mitosis, phosphorylated VCIP135 is inactive and may not be able to bind p97."
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