Werner syndrome protein directly binds to the AAA ATPase p97/VCP in an ATP-dependent fashion

Indig, Fred E.; Partridge, Juneth J.; Kobbe, Cayetano von; Aladjem, Mirit I.; Latterich, Martin; Bohr, Vilhelm A.

doi:10.1016/j.jsb.2003.11.009

Cited by 41 publications

(37 citation statements)

References 45 publications

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“…Furthermore, we have mapped the site of phosphorylation as Ser 784 . These findings suggest a role of VCP in the cellular response to DNA damage and are consistent with previously published interactions of VCP with BRCA1 and the WRN helicase (18)(19)(20).…”

Section: Discussionsupporting

confidence: 92%

“…Because VCP can participate in several macromolecular complexes and can act as a chaperone of many proteins, it is involved in many unrelated cellular activities, such as membrane fusion, cell cycle regulation, stress response, programmed cell death, B-and T-cell activation, transcriptional regulation, endoplasmic reticulum (ER)-associated degradation, and protein degradation (13). VCP has also been proposed to play a role in the DNA damage response, because it can associate with the Werner syndrome protein, a member of the RecQ helicase family (18,19), as well as with BRCA1 (20). However, its precise role in the response of cells to DNA damage is still obscure.…”

Section: Introductionmentioning

confidence: 99%

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Valosin-Containing Protein Phosphorylation at Ser784 in Response to DNA Damage

et al. 2005

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The response of eukaryotic cells to DNA damage includes the activation of phosphatidylinositol-3 kinase-related kinases (PIKK), such as ATM, ATR, and DNA-dependent protein kinase (DNA-PK). These three kinases have very similar substrate specificities in vitro, but in vivo, their substrates overlap only partially. Several in vivo substrates of ATM and ATR have been identified and almost all of them are involved in DNA damageinduced cell cycle arrest and/or apoptosis. In contrast, few in vivo substrates of DNA-PK have been identified. These include histone H2AX and DNA-PK itself. We identify here valosin-containing protein (VCP) as a novel substrate of DNA-PK and other PIKK family members. VCP is phosphorylated at Ser 784 within its COOH terminus, a region previously shown to target VCP to specific intracellular compartments. Furthermore, VCP phosphorylated at Ser 784 accumulated at sites of DNA double-strand breaks (DSBs). VCP is a protein chaperone that unfolds and translocates proteins. Its phosphorylation in response to DNA damage and its recruitment to sites of DNA DSBs could indicate a role of VCP in DNA repair. (Cancer Res 2005; 65(17): 7533-40)

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Section: Discussionsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Valosin-Containing Protein Phosphorylation at Ser784 in Response to DNA Damage

et al. 2005

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“…Given that the CDC-48 UFDϪ1/NPLϪ4 complex is required for embryonic cell division and germline development of adult worms together with its ubiquitous expression (30), we propose a general role in DNA replication of mitotic cells. At present it is unclear how CDC-48 affects replication but recent studies reported that p97 binds to DNA replication factors (31,32). Interestingly, p97 associates with the human Werner RecQ helicase (33) and C. elegans CDC-48.1 with HIM-6, a Bloom Syndrome helicase homolog (34).…”

Section: Resultsmentioning

confidence: 99%

Cell cycle progression requires the CDC-48 ^{UFD−1/NPL−4} complex for efficient DNA replication

Mouysset

Deichsel

Moser

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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Since cdc48 mutants were isolated by the first genetic screens for cell division cycle (cdc) mutants in yeast, the requirement of the chaperone-like ATPase Cdc48/p97 during cell division has remained unclear. Here, we discover an unanticipated function for Caenorhabditis elegans CDC-48 in DNA replication linked to cell cycle control. Our analysis of the CDC-48 UFD؊1/NPL؊4 complex identified a general role in S phase progression of mitotic cells essential for embryonic cell division and germline development of adult worms. These developmental defects result from activation of the DNA replication checkpoint caused by replication stress. Similar to loss of replication licensing factors, DNA content is strongly reduced in worms depleted for CDC-48, UFD-1, and NPL-4. In addition, these worms show decreased DNA synthesis and hypersensitivity toward replication blocking agents. Our findings identified a role for CDC-48 UFD؊1/NPL؊4 in DNA replication, which is important for cell cycle progression and genome stability.ATL-1/ATR ͉ C. elegans ͉ CDC-48/p97 ͉ genome stability M any biological processes including development and cell division are tightly controlled by ubiquitin-mediated protein degradation. A central factor for mobilizing and targeting ubiquitylated substrates to the 26S proteasome is Cdc48/p97 (Cdc48 in yeast, CDC-48 in C. elegans, p97 in mammals), a chaperone-like AAA ATPase (1). Its activity is modulated by alternative adaptor proteins, which determine recruitment and processing of specific substrates. Cdc48/p97 forms a complex with the cofactors Ufd1 and Npl4 that is involved in endoplasmic reticulum (ER)-associated protein degradation (ERAD) (2), membrane fusion and cell cycle progression (3, 4). Temperature sensitive cdc48 mutants have already been isolated by early cdc-screens (5) in Saccharomyces cerevisiae. However, the essential role of Cdc48 during cell cycle progression remained elusive.Meanwhile, different activities of Cdc48/p97 in mitosis have been addressed by several studies. Early observations in yeast and recent findings using Xenopus egg extracts suggested that Cdc48/p97 regulates spindle disassembly during exit from mitosis (6, 7). For example, spindle regulators such as the Polo-like kinase Plx remain attached and probably stabilize the spindle in the absence of p97 Ufd1/Npl4 . However, contradictory evidence exists concerning a specific role of the p97 Ufd1/Npl4 complex in spindle dynamics (8,9). Beside spindle function, p97 together with its Ufd1-Npl4 cofactor is important for nuclear envelope assembly (10). Interestingly, it has been shown that p97 stimulates nucleus reformation after mitosis by extracting and thereby inactivating the mitotic progression kinase Aurora B from chromatin (11). Together, these diverse processes involving Cdc48/p97 suggest the existence of multiple substrates that need to be regulated during mitosis.Recently, we found that the C. elegans Cdc48/p97 homologues CDC-48.1 and CDC-48.2 form an evolutionarily conserved complex with UFD-1 and NPL-4 important for t...

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“…p97/VCP is an essential biochemical component of a wide range of ubiquitin-linked cell biological reactions, including ubiquitin-proteasome system-mediated protein degradation (8), Golgi and endoplasmic reticulum (ER) membrane fusion (1,18), transcription factor activation (19), and DNA repair (10,16). In these processes, p97 acts as a molecular segregase that utilizes ATP-powered conformational changes in the assembly and disassembly of macromolecular machineries (8,11).…”

mentioning

confidence: 99%

Hereditary Inclusion Body Myopathy-Linked p97/VCP Mutations in the NH₂ Domain and the D1 Ring Modulate p97/VCP ATPase Activity and D2 Ring Conformation

Halawani

LeBlanc

Rouiller

et al. 2009

Molecular and Cellular Biology

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105

131

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Hereditary inclusion body myopathy associated with early-onset Paget disease of bone and frontotemporal dementia (hIBMPFTD) is a degenerative disorder caused by single substitutions in highly conserved residues of p97/VCP. All mutations identified thus far cluster within the NH 2 domain or the D1 ring, which are both required for communicating conformational changes to adaptor protein complexes. In this study, biochemical approaches were used to identify the consequences of the mutations R155P and A232E on p97/VCP structure. Assessment of p97/VCP oligomerization revealed that p97 R155P and p97 A232E formed hexameric ring-shaped structures of ϳ600 kDa. p97 R155P and p97 A232E exhibited an ϳ3-fold increase in ATPase activity compared to wild-type p97 (p97 WT ) and displayed increased sensitivity to heat-induced upregulation of ATPase activity. Protein fluorescence analysis provided evidence for conformational differences in the D2 rings of both hIBMPFTD mutants. Furthermore, both mutations increased the proteolytic susceptibility of the D2 ring. The solution structures of all p97/VCP proteins revealed a didispersed distribution of a predominant hexameric population and a minor population of large-diameter complexes. ATP binding significantly increased the abundance of large-diameter complexes for p97 R155P and p97 A232E , but not p97 WT or the ATP-binding mutant p97 K524A . Therefore, we propose that hIBMPFTD p97/VCP mutants p97 R155P and p97 A232E possess structural defects that may compromise the mechanism of p97/VCP activity within large multiprotein complexes.The ubiquitous valosin-containing protein (p97/VCP) is a prominent member of the highly conserved AAA ϩ (ATPases associated with diverse cellular activities) proteins, which are known for their oligomeric structure and chaperone-like activities. p97/VCP is an essential biochemical component of a wide range of ubiquitin-linked cell biological reactions, including ubiquitin-proteasome system-mediated protein degradation (8), Golgi and endoplasmic reticulum (ER) membrane fusion (1, 18), transcription factor activation (19), and DNA repair (10, 16). In these processes, p97 acts as a molecular segregase that utilizes ATP-powered conformational changes in the assembly and disassembly of macromolecular machineries (8, 11).p97 ATPase activity is contingent on the assembly of an inherently stable hexamer (24) comprised of two highly homologous D1 and D2 nucleotide-binding rings and regulatory NH 2 -and COOH-terminal domains (4). ATP hydrolysis in the D2 ring mediates p97 major ATPase activity, while the D1 ring is involved in the regulation of p97 hexamerization (24). Recently, p97 has been linked to a severe degenerative disorder identified as hereditary inclusion body myopathy associated with early-onset Paget disease of bone and frontotemporal dementia (hIBMPFTD). The pathogenesis of hIBMPFTD is attributed to autosomal-dominant single-amino-acid substitutions in highly conserved residues within the p97 NH 2 domain and D1 ring (28). Patients present with hallma...

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Werner syndrome protein directly binds to the AAA ATPase p97/VCP in an ATP-dependent fashion

Cited by 41 publications

References 45 publications

Valosin-Containing Protein Phosphorylation at Ser784 in Response to DNA Damage

Valosin-Containing Protein Phosphorylation at Ser784 in Response to DNA Damage

Cell cycle progression requires the CDC-48 ^{UFD−1/NPL−4} complex for efficient DNA replication

Hereditary Inclusion Body Myopathy-Linked p97/VCP Mutations in the NH₂ Domain and the D1 Ring Modulate p97/VCP ATPase Activity and D2 Ring Conformation

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Werner syndrome protein directly binds to the AAA ATPase p97/VCP in an ATP-dependent fashion

Cited by 41 publications

References 45 publications

Valosin-Containing Protein Phosphorylation at Ser784 in Response to DNA Damage

Valosin-Containing Protein Phosphorylation at Ser784 in Response to DNA Damage

Cell cycle progression requires the CDC-48 UFD−1/NPL−4 complex for efficient DNA replication

Hereditary Inclusion Body Myopathy-Linked p97/VCP Mutations in the NH2 Domain and the D1 Ring Modulate p97/VCP ATPase Activity and D2 Ring Conformation

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Resources

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Cell cycle progression requires the CDC-48 ^{UFD−1/NPL−4} complex for efficient DNA replication

Hereditary Inclusion Body Myopathy-Linked p97/VCP Mutations in the NH₂ Domain and the D1 Ring Modulate p97/VCP ATPase Activity and D2 Ring Conformation