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VCPIP1 binds VCP. 82 / 82
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sparser
"Here, we use cryo-EM to characterize both VCP-VCPIP1 and VCP-VCPIP1-p47 complexes."
40877265
sparser
"To understand how VCP engages with VCPIP1 and p47, we determined high resolution cryogenic electron microscopy (cryo-EM) structures of both VCP-VCPIP1 and VCP-VCPIP1-p47 complexes capturing interaction states at residue resolution despite the high conformational flexibility."
40877265
sparser
"To put the VCP-VCPIP1 structure in context with the critical VCP adapter p47, we determined the structure of a VCP-VCPIP1-p47 ternary complex, which reveals that the p47 UBX domain outcompetes the VCPIP1 UBX domain on the VCP N-domain, which may suggest dynamic competition for N-domain binding, as seen with other VCP cofactors xref , xref , xref and in other VCP processes."
40877265
sparser
"In this work, we demonstrate how VCPIP1 bivalently binds to VCP via the UBX domain and the newly characterized VCPID and how these domains along with its DUB activity are crucial for Golgi assembly."
40877265
biogrid
No evidence text available
23500464
biogrid
No evidence text available
23500464
sparser
"VCP interacts with VCPIP1 through multivalent interactions."
40877265
sparser
"To reconstitute the VCP-VCPIP1 complex for structural studies, we co-expressed and purified full-length VCPIP1 and VCP (Supplementary Fig.  xref , see Fig.  xref for schematics of domain organization indicating domain boundaries) from Expi293 cells with no addition of ATP or ATP analogs (see “Methods”)."
40877265
sparser
"We obtained a consensus reconstruction of the VCP-VCPIP1 complex at an overall resolution of 2.3 Å (see “Methods” and Supplementary Figs.  xref , xref for details)."
40877265
sparser
"Taken together, our cryo-EM structure of shows VCPIP1 bound to VCP in a 1:2 stoichiometry, although we cannot fully exclude the possibility that additional VCPIP1 could engage through the N-domain only."
40877265
biogrid
No evidence text available
29540532
biogrid
No evidence text available
26389662
sparser
"Since previous studies have shown that bound nucleotide influences the conformation of the N-domain xref , xref , xref , we asked whether it may also affect binding of VCPIP1 to VCP."
40877265
sparser
"These data suggest that for VCPIP1 binding to VCP, locking the N-domains in either the up- or down- positions presents an energetic barrier which may be due to the conformation of VCPIP1 bound VCP being slightly different to the locked states."
40877265
sparser
"After initial rounds of 3D classification, we were again able to observe density in the two binding regions on VCP, similar to what we observed for the VCP-VCPIP1 complex (see “Methods” and Supplementary Figs.  xref , xref for details)."
40877265
sparser
"The binding mode of the three VCPIDs bound to the D2 dimer is indistinguishable from the binding mode observed in the VCP-VCPIP1 complex, with the stalk regions connected to each VCPID clearly visible (Fig.  xref )."
40877265
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"VCIP135 binds to the p97/p47/syntaxin5 complex and promotes its dissociation via p97 catalysed ATP hydrolysis ( Uchiyama et al., 2002 )."
14643202
sparser
"VCPIP1 interactions with VCP and catalytic activity are required for effect on Golgi."
40877265
sparser
"Next, we asked whether the interaction of VCPIP1 with VCP is necessary for the previously reported role of VCPIP1 in Golgi reassembly."
40877265
reach
"We determine a ∼3Å cryo-EM structure of the VCP-VCPIP1 complex and find up to 3 VCPIP1 protomers interact with the VCP hexamer."
39314330
sparser
"Our structure of the VCP-VCPIP1 complex reveals that, unlike other structurally characterized VCP-adapters that show a single point of contact, VCPIP1 forms two distinct interfaces with VCP."
40877265
sparser
"During the preparation of our manuscript, two studies by Liao et al. xref and Vostal et al. xref also reported on VCP-VCPIP1 complexes describing overall similar arrangements of the complex and reporting several overlapping findings that mutually reinforce the work."
40877265
sparser
"Although exogenous ATPs were not used for structural studies, VCP N-domains were seen in the up and down conformation for the VCP-VCPIP1 complex and further refinement with six-fold symmetry was done in both conformations, but there was substantially less UBX domain density bound in the up-position compared to the down-position (see intermediates in Supplementary Fig.  xref )."
40877265
sparser
"This model is further supported by sensitivity of VCPIP1 binding to VCP to nucleotide state (see Supplementary Fig.  xref )."
40877265
biogrid
No evidence text available
21811234
sparser
"We determine ∼3.3 Å cryogenic electron microscopy structures of VCP-VCPIP1 complexes in the absence of added nucleotide or the presence of an ATP analog."
40085114
reach
"Additionally, the binding of VCIP135 to p97 was reported to cause no change in its deubiquitinating activity (9)."
38072049
hprd
No evidence text available
12473691
reach
"To date, there are no lines of evidence indicating that VCIP135 requires its binding to p97 for Golgi and ER biogenesis.In this study, we investigated the binding interactions between VCIP135 and p97 and found that VCIP135 has two distinct p97-binding sites; that is, its N- and C-terminal p97-binding sites interact with the C- and N-terminal regions of p97, respectively."
38072049
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"Regarding the p97-binding sites in its C-terminal half, only VCIP135(903-1053) bound to p97, but the other fragments did not (Fig. 1D)."
38072049
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"These results strongly suggest that VCIP135 can associate with p97 via two distinct interactions."
38072049
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"VCIP135(1-743)wt bound to p97 (top panel, lane 2), whereas VCIP135(1-743)(L133S) showed almost no binding to p97 (top panel, lane 3)."
38072049
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"The VCIP135 mutants (L133S) and (F1024L/L1031P), in which only one of the two p97-binding sites was intact, still bound to p97, although their binding affinities were decreased compared with that of VCIP135wt (Fig. 5A, lanes 2–4; Fig. S1D)."
38072049
reach
"In other words, these results strongly support the fact that each of the two distinct binding interactions between VCIP135 and p97 is sufficient for VCIP135-p97 complex formation.We also investigated the nucleotide dependency of the two interactions between VCIP135 and p97 using these full-length VCIP135 mutants."
38072049
sparser
"In conclusion, we report six new models of VCP bound to VCPIP1 and in a ternary complex of VCP-VCPIP1-p47."
40877265
reach
"These results indicate that the binding of ATP to p97 enhances both of these interactions between VCIP135 and p97; that is, these two interactions in the complex may be synchronously controlled by the nucleotide state of p97."
38072049
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"The two interactions between VCIP135 and p97 are required for Golgi and ER biogenesis in vivo."
38072049
sparser
"The high resolution of our structures, along with a heterogeneity and dynamics aware processing compared to similar studies xref , xref enabled us to uniquely identify a high degree of dynamics and flexibility in VCP-VCPIP1 and VCP-VCPIP1-p47 complexes."
40877265
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"We previously reported that both VCIP135 and p97, which form a complex in the cytosol, bind to Golgi and ER membranes, and function in Golgi and ER biogenesis (6, 8)."
38072049
reach
"As p97 is an ATPase, we first analyzed the effects of nucleotides on the binding between p97 and VCIP135."
38072049
sparser
"Investigators show that VCPIP1 interacts with VCP and other genes associated with the reassembly of the Golgi apparatus and the endoplasmic reticulum during mitosis [ xref , xref ]."
39796135
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"Figure 1A shows that the addition of AMP-PNP, a nonhydrolyzable ATP analogue, increased the binding of VCIP135 to p97 (lane 4; Fig. S1A), suggesting that ATP binding enhances the association between p97 and VCIP135."
38072049
reach
"VCIP135(1-390) bound to p97, whereas VCIP135(361-743) did not."
38072049
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"However, it remains unclear whether the interactions between VCIP135 and p97 are required for their biogenesis."
38072049
reach
"These in vivo results strongly suggest that both of the two interactions between VCIP135 and p97 are required for Golgi biogenesis.We also previously reported that both VCIP135 and p97 bind to ER membranes and function in ER network formation (8, 9)."
38072049
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"We therefore investigated the roles of the interactions between VCIP135 and p97 in ER network formation in living cells."
38072049
reach
"Third, these two binding interactions in the complex are synchronously controlled by the nucleotide state of p97 (Fig. 5, C and D), which leads to the nucleotide-dependent association between full-length VCIP135 and p97 (Fig. 1A)."
38072049
reach
"Golgi WAC interacts with the deubiquitinase VCIP135, which binds VCP and p97."
26687599
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"However, the ATPase assay revealed that VCPIP1 binding to P97/VCP only slightly increases its ATPase activity (Figure 1E).2."
39234822
reach
"Finally, each of the two binding interactions between VCIP135 and p97 is sufficient for their complex formation."
38072049
reach
"The phosphorylation of Ser-130 might disrupt the binding of p97 to the N-terminal binding site in VCIP135.In the present study, we clarified that VCIP135 and p97 form a complex through two distinct interactions and that either one of the interactions is sufficient for this complex formation."
38072049
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"This indicates that the association between p97 and VCIP135 is necessary for Golgi and ER biogenesis."
38072049
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"Interestingly, these morphological changes were not rescued by the expression of VCIP135 mutants with only one of the two p97 binding sites, indicating that these two distinct interactions between VCIP135 and p97 are essential for Golgi and ER biogenesis."
38072049
reach
"We also showed that the binding of VCIP135 to p97 does not change its deubiquitinating activity (9)."
38072049
reach
"Taking these facts altogether, it is plausible that these binding interactions of VCIP135 with p97 may be required for the ubiquitin-independent functions of VCIP135, but not for its ubiquitin-dependent function.Finally, we will discuss the functional significance of these two binding interactions in the VCIP135-p97 complex."
38072049
biogrid
No evidence text available
19615732
reach
"The tethering complex is generally thought to be dissociated and then reused after membrane fusion, and hence VCIP135 might be a candidate recycling factor of the tethering complex.Another important point is that VCIP135 can associate with p97 by binding to either its N- or C-terminal region."
38072049
biogrid
No evidence text available
29599191
sparser
"We determine a ∼3Å cryo-EM structure of the VCP-VCPIP1 complex and find up to 3 VCPIP1 protomers interact with the VCP hexamer."
39314330
biogrid
No evidence text available
24163436
reach
"Conversely, VCPIP1 bound P97/VCP hexamer favors the binding of P47, and thus the intact SNARE complex, promoting Golgi membrane fusion."
39234822
reach
"28 However, the molecular mechanism of VCPIP1 interacting with P97/VCP and the precise roles of the VCPIP1‐P97/VCP complex in Golgi membrane fusion in late mitosis is still elusive.In this study, we first analyzed the interactions between P97/VCP and VCPIP1 fragments and found out that VCPIP1 can associate with P97/VCP in vitro through multiple binding sites, including the N‐terminal OTU domain, the middle UFD1 domain, and the C‐terminal UBL‐UFD2 domain."
39234822
reach
"The results reveal that, in the absence of substrates, the binding of VCPIP1 to P97/VCP led to a marginal increase in its activity (Figure 1E).2.2 Cryo-EM Structure of VCPIP1-P97/VCP Complex."
39234822
reach
"Albeit with the relatively lower and heterogeneous resolution of VCPIP1, the regions of VCPIP1 that directly contact P97/VCP could be readily and confidently modeled.Expectedly, the P97/VCP in the P97/VCP‐VCPIP1 complex was folded into the canonical homo‐hexamer as previously studied (Figure 2A,C)."
39234822
reach
"Unexpectedly, three VCPIP1 molecules directly bind to the C‐terminal D2 ATPase domain of the P97/VCP, resulting in the formation of P97/VCP‐VCPIP1 complex with a stoichiometry of 2:1 (Figure 2B)."
39234822
reach
"Moreover, we successfully determined the Cryo‐EM structure of the full‐length P97/VCP‐VCPIP1 complex, which reveals that three individual VCPIP1 molecules directly bind to the C‐terminal D2 domains of the P97/VCP hexamer through their UFD1 domains."
39234822
reach
"Overall, our study not only reveals the unexpected interaction and organization of human P97/VCP‐VCPIP1 complex, but also provides crucial insights into the mechanism of VCPIP1‐P97/VCP mediated Golgi biogenesis.2 Results."
39234822
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"To validate the direct interaction between VCPIP1 and P97/VCP in vitro, we first purified the full‐length proteins and analyzed their interactions by size exclusion chromatography."
39234822
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"The obtained result showed that the VCPIP1 could directly bind to P97/VCP by migrating together with P97/VCP on the size exclusion column (Figure 1B)."
39234822
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"Since only one 3D reconstruction was obtained and could be refined to high resolution from our cryo‐EM studies, we favored it as the stable and dominant conformation adopted by the full‐length P97/VCP‐VCPIP1 complex (Figure S1C, Supporting Information)."
39234822
reach
"We concluded that this UFD1‐mediated VCPIP1 interaction with P97/VCP is critical for their function in Golgi reassembly."
39234822
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"Whether the interaction between P97/VCP and VCPIP1 is directly regulated by CDK1 or other mitotic kinases needs further exploration."
39234822
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"34 Considering the key role in contact with VCP by VCPIP1 Y623 and F638, which are the neighboring residues of K632 and K646, UFD1 ubiquitination might have a direct effect on their interaction and thus regulate Golgi membrane dynamics.In conclusion, our findings uncover the high‐resolution structure of the P97/VCP‐VCPIP1 complex and reveal the unique and unexpected binding modes between the ATP‐driven substrate unfoldase P97/VCP and the deubiquitinase VCPIP1."
39234822
reach
"The resulting structural model indicates that VCPIP1 binds to P97/VCP through three major interfaces, with both the N‐terminal and C‐terminal regions of P97/VCP required for VCPIP1 binding (Figure 4A)."
39234822
reach
"Previous studies have proved that VCPIP1 interaction with P97/VCP synergistically facilitates the post‐mitotic Golgi vesicle fusion by regulating the priming of syntaxin‐5 containing SNARE complex through the P97/VCP adaptor protein P47 during the cell cycle."
39234822
reach
"These biochemical assays further demonstrated that VCPIP1 contacts the D2 domain of P97/VCP through its UFD1 domain, inducing a conformational change in P97/VCP to favor its binding to the SNARE substrate, thereby regulating the disassembly of the SNARE complex.To further investigate the functional relevance of the P97/VCP‐VCPIP1 complex in post‐mitotic Golgi reassembly, we generated a VCPIP1‐knockout HeLa cell line using CRISPR‐Cas9 technology."
39234822
reach
"In this study, our structural and functional studies of the P97/VCP‐VCPIP1 complex provide key insights into the organization and membrane fusion mechanism of the underappreciated P97/VCP‐VCPIP1 complex.VCPIP1 was initially identified as an essential factor for P97/VCP/p47‐mediated Golgi and ER assembly (15)."
39234822
signor
"Golgi biogenesis requires two distinct p97ATPase-mediated membrane fusion, the p97/p47 and p97/p37 pathways. |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
reach
"P97 binds a variety of proteins with various functions, for example VCIP135 (valosin-containing protein (p97)/p47 complex-interacting protein p135) [3] and p47."
15498563
sparser
"Previous studies have demonstrated that VCP, in complex with VCPIP1, is necessary for cisternal regrowth xref , and that p47 may serve to recruit the VCP-VCPIP1 complex to the substrate xref , xref , xref ."
40877265
biogrid
No evidence text available
36300783
biogrid
No evidence text available
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