title: Valosin-containing Protein (VCP)/p97 Segregase Mediates Proteolytic Processing of Cockayne Syndrome Group B (CSB) in Damaged Chromatin.
abstract: Cockayne syndrome group A and B (CSB) proteins act in transcription-coupled repair, a subpathway of nucleotide excision repair. Here we demonstrate that valosin-containing protein (VCP)/p97 segregase functions in ultraviolet radiation (UVR)-induced ubiquitin-mediated CSB degradation. We show that VCP/p97 inhibition and siRNA-mediated ablation of VCP/p97 and its cofactors UFD1 and UBXD7 impair CSB degradation. VCP/p97 inhibition also results in the accumulation of CSB in chromatin. Moreover, VCP/p97 interacts with both native and ubiquitin-conjugated forms of CSB. The localized cellular UVR exposures lead to VCP/p97 accumulation at DNA damage spots, forming distinct UVR-induced foci. However, manifestation of VCP/p97 foci is independent of CSB and UBXD7. Furthermore, VCP/p97 and UBXD7 associate with the Cockayne syndrome group A-DDB1-Cul4A complex, an E3 ligase responsible for CSB ubiquitination. Compromising proteasome and VCP/p97 function allows accumulation of both native and ubiquitinated CSB and results in an increase of UBXD7, proteasomal RPN2, and Sug1 in the chromatin compartment. Surprisingly, both biochemical inhibition and genetic defect of VCP/p97 enhance the recovery of RNA synthesis following UVR, whereas both VCP/p97 and proteasome inhibitions decrease cell viability. Our findings reveal a new role of VCP/p97 segregase in the timely processing of ubiquitinated CSB from damaged chromatin.
authors.affiliation: From the Departments of Radiology and.
authors.forename: Jinshan
authors.initials: J
authors.lastname: He
authors.affiliation: From the Departments of Radiology and zhu.49@osu.edu.
authors.forename: Qianzheng
authors.initials: Q
authors.lastname: Zhu
authors.affiliation: From the Departments of Radiology and.
authors.forename: Gulzar
authors.initials: G
authors.lastname: Wani
authors.affiliation: From the Departments of Radiology and.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Sharma
authors.affiliation: From the Departments of Radiology and Molecular and Cellular Biochemistry and James Cancer Hospital and Solove Research Institute, Ohio State University, Columbus, Ohio 43210 wani.2@osu.edu.
authors.forename: Altaf A
authors.initials: AA
authors.lastname: Wani
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D044842:Cullin Proteins / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D045643:DNA Repair Enzymes / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins / Q000235:genetics / Q000378:metabolism; D000075223:Poly-ADP-Ribose Binding Proteins; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D059748:Proteolysis; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination / Q000235:genetics / Q000528:radiation effects; D014466:Ultraviolet Rays / Q000009:adverse effects*; D000074405:Valosin Containing Protein
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; C114234:CUL4A protein, human; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002843:Chromatin; D044842:Cullin Proteins; C091435:DDB1 protein, human; D004268:DNA-Binding Proteins; C095472:ERCC8 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; C092282:PSMC5 protein, human; D000075223:Poly-ADP-Ribose Binding Proteins; D011506:Proteins; D014157:Transcription Factors; C531895:UBXN7 protein, human; C491497:UFD1 protein, human; D006602:Hexosyltransferases; C517590:RPN2 protein, human; D046988:Proteasome Endopeptidase Complex; D000251:Adenosine Triphosphatases; D000074183:ATPases Associated with Diverse Cellular Activities; D004265:DNA Helicases; C078673:ERCC6 protein, human; C000614178:VCP protein, human; D000074405:Valosin Containing Protein; D045643:DNA Repair Enzymes
keywords: Cockayne syndrome group B protein; DNA damage; DNA repair; nucleotide excision repair; protein degradation; transcription-coupled repair; ubiquitin; valosin-containing protein
references.citation: Mol Cell Biol. 2000 Oct;20(20):7643-53
references.pmid: 11003660
references.citation: Nucleic Acids Res. 2005;33(13):4023-34
references.pmid: 16030353
references.citation: Mol Biol Cell. 2001 Dec;12(12):4114-28
references.pmid: 11739805
references.citation: Nature. 2001 Dec 6;414(6864):652-6
references.pmid: 11740563
references.citation: Nat Rev Mol Cell Biol. 2002 Jan;3(1):21-9
references.pmid: 11823795
references.citation: Nat Cell Biol. 2002 Feb;4(2):134-9
references.pmid: 11813000
references.citation: EMBO J. 2002 Feb 15;21(4):615-21
references.pmid: 11847109
references.citation: DNA Repair (Amst). 2011 Jul 15;10(7):697-713
references.pmid: 21571596
references.citation: Cancer Sci. 2011 Oct;102(10):1840-7
references.pmid: 21756275
references.citation: Mol Cell. 2005 Oct 28;20(2):187-98
references.pmid: 16246722
references.citation: J Biol Chem. 2006 May 12;281(19):13404-11
references.pmid: 16527807
references.citation: Genes Dev. 2006 Jun 1;20(11):1429-34
references.pmid: 16751180
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 20;103(25):9613-8
references.pmid: 16772382
references.citation: Mol Cell. 2006 Aug;23(4):471-82
references.pmid: 16916636
references.citation: J Struct Biol. 2006 Oct;156(1):29-40
references.pmid: 16529947
references.citation: Mol Cell. 2007 Aug 17;27(4):585-95
references.pmid: 17707230
references.citation: Mol Cell. 2007 Nov 9;28(3):386-97
references.pmid: 17996703
references.citation: Nature. 2007 Dec 20;450(7173):1258-62
references.pmid: 18097415
references.citation: Cell. 2008 Sep 5;134(5):804-16
references.pmid: 18775313
references.citation: J Biol Chem. 2008 Oct 31;283(44):30289-99
references.pmid: 18715868
references.citation: Mol Cell. 2011 Jan 7;41(1):82-92
references.pmid: 21211725
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):4834-9
references.pmid: 21383145
references.citation: Trends Biochem Sci. 2011 Oct;36(10):515-23
references.pmid: 21741246
references.citation: Nat Cell Biol. 2011 Nov;13(11):1376-82
references.pmid: 22020440
references.citation: Nat Struct Mol Biol. 2011 Dec;18(12):1345-50
references.pmid: 22120668
references.citation: Nat Cell Biol. 2012 Feb;14(2):117-23
references.pmid: 22298039
references.citation: Nat Struct Mol Biol. 2012 May;19(5):511-6, S1
references.pmid: 22466964
references.citation: Nat Genet. 2012 May;44(5):593-7
references.pmid: 22466612
references.citation: Nat Genet. 2012 May;44(5):598-602
references.pmid: 22466611
references.citation: BMC Biol. 2012;10:48
references.pmid: 22694940
references.citation: J Biol Chem. 2012 Oct 12;287(42):35118-26
references.pmid: 22902626
references.citation: PLoS Genet. 2013 Apr;9(4):e1003407
references.pmid: 23637612
references.citation: Cell Cycle. 2014;13(1):106-14
references.pmid: 24196443
references.citation: Nat Commun. 2014;5:3695
references.pmid: 24770583
references.citation: J Biol Chem. 2014 Sep 26;289(39):27278-89
references.pmid: 25118285
references.citation: Proc Natl Acad Sci U S A. 2014 Oct 7;111(40):14454-9
references.pmid: 25249633
references.citation: Cell Cycle. 2015;14(9):1413-25
references.pmid: 25894431
references.citation: J Cell Sci. 2003 Feb 1;116(Pt 3):447-51
references.pmid: 12508106
references.citation: Cell. 2003 May 2;113(3):357-67
references.pmid: 12732143
references.citation: J Biol Chem. 2003 Nov 21;278(47):46906-10
references.pmid: 12944386
references.citation: Nature. 2004 Apr 1;428(6982):1 p following 486
references.pmid: 15058298
references.citation: J Cell Biol. 2004 Jul 5;166(1):27-36
references.pmid: 15226310
references.citation: Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11586-90
references.pmid: 8876179
references.citation: Curr Top Microbiol Immunol. 1997;221:47-70
references.pmid: 8979440
references.citation: EMBO J. 1997 Oct 1;16(19):5955-65
references.pmid: 9312053
references.citation: Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11205-9
references.pmid: 9326587
references.citation: J Biol Chem. 1998 May 8;273(19):11844-51
references.pmid: 9565609
references.citation: Genes Dev. 1999 Apr 1;13(7):768-85
references.pmid: 10197977
references.citation: Mol Cell. 2001 Jul;8(1):213-24
references.pmid: 11511374