Ataxia telangiectasia-mutated damage-signaling kinase- and proteasome-dependent destruction of Mre11-Rad50-Nbs1 subunits in simian virus 40-infected primate cells

Xiaorong Zhao, Ramiro J. Madden-Fuentes, Becky X. Lou, James M. Pipas, Jeannine Gerhardt, Christopher J. Rigell, Ellen Fanning

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Although the mechanism of simian virus 40 (SV40) DNA replication has been extensively investigated with cell extracts, viral DNA replication in productively infected cells utilizes additional viral and host functions whose interplay remains poorly understood. We show here that in SV40-infected primate cells, the activated ataxia telangiectasia-mutated (ATM) damage-signaling kinase, γ-H2AX, and Mre11-Rad50-Nbs1 (MRN) assemble with T antigen and other viral DNA replication proteins in large nuclear foci. During infection, steady-state levels of MRN subunits decline, although the corresponding mRNA levels remain unchanged. A proteasome inhibitor stabilizes the MRN complex, suggesting that MRN may undergo proteasome-dependent degradation. Analysis of mutant T antigens with disrupted binding to the ubiquitin ligase CUL7 revealed that MRN subunits are stable in cells infected with mutant virus or transfected with mutant viral DNA, implicating CUL7 association with T antigen in MRN proteolysis. The mutant genomes produce fewer virus progeny than the wild type, suggesting that T antigen-CUL7-directed proteolysis facilitates virus propagation. Use of a specific ATM kinase inhibitor showed that ATM kinase signaling is a prerequisite for proteasome-dependent degradation of MRN subunits as well as for the localization of T antigen and damage-signaling proteins to viral replication foci and optimal viral DNA replication. Taken together, the results indicate that SV40 infection manipulates host DNA damage-signaling to reprogram the cell for viral replication, perhaps through mechanisms related to host recovery from DNA damage.

Original languageEnglish (US)
Pages (from-to)5316-5328
Number of pages13
JournalJournal of Virology
Volume82
Issue number11
DOIs
StatePublished - Jun 2008
Externally publishedYes

Fingerprint

Simian virus 40
Ataxia Telangiectasia
Viral Tumor Antigens
proteasome endopeptidase complex
Proteasome Endopeptidase Complex
Primates
Viral DNA
DNA replication
phosphotransferases (kinases)
DNA Replication
Phosphotransferases
antigens
mutants
virus replication
Viruses
proteolysis
DNA damage
viruses
Proteolysis
DNA Damage

ASJC Scopus subject areas

  • Immunology

Cite this

Ataxia telangiectasia-mutated damage-signaling kinase- and proteasome-dependent destruction of Mre11-Rad50-Nbs1 subunits in simian virus 40-infected primate cells. / Zhao, Xiaorong; Madden-Fuentes, Ramiro J.; Lou, Becky X.; Pipas, James M.; Gerhardt, Jeannine; Rigell, Christopher J.; Fanning, Ellen.

In: Journal of Virology, Vol. 82, No. 11, 06.2008, p. 5316-5328.

Research output: Contribution to journalArticle

Zhao, Xiaorong ; Madden-Fuentes, Ramiro J. ; Lou, Becky X. ; Pipas, James M. ; Gerhardt, Jeannine ; Rigell, Christopher J. ; Fanning, Ellen. / Ataxia telangiectasia-mutated damage-signaling kinase- and proteasome-dependent destruction of Mre11-Rad50-Nbs1 subunits in simian virus 40-infected primate cells. In: Journal of Virology. 2008 ; Vol. 82, No. 11. pp. 5316-5328.
@article{5a87c8271352489ba796167d94d0d93d,
title = "Ataxia telangiectasia-mutated damage-signaling kinase- and proteasome-dependent destruction of Mre11-Rad50-Nbs1 subunits in simian virus 40-infected primate cells",
abstract = "Although the mechanism of simian virus 40 (SV40) DNA replication has been extensively investigated with cell extracts, viral DNA replication in productively infected cells utilizes additional viral and host functions whose interplay remains poorly understood. We show here that in SV40-infected primate cells, the activated ataxia telangiectasia-mutated (ATM) damage-signaling kinase, γ-H2AX, and Mre11-Rad50-Nbs1 (MRN) assemble with T antigen and other viral DNA replication proteins in large nuclear foci. During infection, steady-state levels of MRN subunits decline, although the corresponding mRNA levels remain unchanged. A proteasome inhibitor stabilizes the MRN complex, suggesting that MRN may undergo proteasome-dependent degradation. Analysis of mutant T antigens with disrupted binding to the ubiquitin ligase CUL7 revealed that MRN subunits are stable in cells infected with mutant virus or transfected with mutant viral DNA, implicating CUL7 association with T antigen in MRN proteolysis. The mutant genomes produce fewer virus progeny than the wild type, suggesting that T antigen-CUL7-directed proteolysis facilitates virus propagation. Use of a specific ATM kinase inhibitor showed that ATM kinase signaling is a prerequisite for proteasome-dependent degradation of MRN subunits as well as for the localization of T antigen and damage-signaling proteins to viral replication foci and optimal viral DNA replication. Taken together, the results indicate that SV40 infection manipulates host DNA damage-signaling to reprogram the cell for viral replication, perhaps through mechanisms related to host recovery from DNA damage.",
author = "Xiaorong Zhao and Madden-Fuentes, {Ramiro J.} and Lou, {Becky X.} and Pipas, {James M.} and Jeannine Gerhardt and Rigell, {Christopher J.} and Ellen Fanning",
year = "2008",
month = "6",
doi = "10.1128/JVI.02677-07",
language = "English (US)",
volume = "82",
pages = "5316--5328",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "11",

}

TY - JOUR

T1 - Ataxia telangiectasia-mutated damage-signaling kinase- and proteasome-dependent destruction of Mre11-Rad50-Nbs1 subunits in simian virus 40-infected primate cells

AU - Zhao, Xiaorong

AU - Madden-Fuentes, Ramiro J.

AU - Lou, Becky X.

AU - Pipas, James M.

AU - Gerhardt, Jeannine

AU - Rigell, Christopher J.

AU - Fanning, Ellen

PY - 2008/6

Y1 - 2008/6

N2 - Although the mechanism of simian virus 40 (SV40) DNA replication has been extensively investigated with cell extracts, viral DNA replication in productively infected cells utilizes additional viral and host functions whose interplay remains poorly understood. We show here that in SV40-infected primate cells, the activated ataxia telangiectasia-mutated (ATM) damage-signaling kinase, γ-H2AX, and Mre11-Rad50-Nbs1 (MRN) assemble with T antigen and other viral DNA replication proteins in large nuclear foci. During infection, steady-state levels of MRN subunits decline, although the corresponding mRNA levels remain unchanged. A proteasome inhibitor stabilizes the MRN complex, suggesting that MRN may undergo proteasome-dependent degradation. Analysis of mutant T antigens with disrupted binding to the ubiquitin ligase CUL7 revealed that MRN subunits are stable in cells infected with mutant virus or transfected with mutant viral DNA, implicating CUL7 association with T antigen in MRN proteolysis. The mutant genomes produce fewer virus progeny than the wild type, suggesting that T antigen-CUL7-directed proteolysis facilitates virus propagation. Use of a specific ATM kinase inhibitor showed that ATM kinase signaling is a prerequisite for proteasome-dependent degradation of MRN subunits as well as for the localization of T antigen and damage-signaling proteins to viral replication foci and optimal viral DNA replication. Taken together, the results indicate that SV40 infection manipulates host DNA damage-signaling to reprogram the cell for viral replication, perhaps through mechanisms related to host recovery from DNA damage.

AB - Although the mechanism of simian virus 40 (SV40) DNA replication has been extensively investigated with cell extracts, viral DNA replication in productively infected cells utilizes additional viral and host functions whose interplay remains poorly understood. We show here that in SV40-infected primate cells, the activated ataxia telangiectasia-mutated (ATM) damage-signaling kinase, γ-H2AX, and Mre11-Rad50-Nbs1 (MRN) assemble with T antigen and other viral DNA replication proteins in large nuclear foci. During infection, steady-state levels of MRN subunits decline, although the corresponding mRNA levels remain unchanged. A proteasome inhibitor stabilizes the MRN complex, suggesting that MRN may undergo proteasome-dependent degradation. Analysis of mutant T antigens with disrupted binding to the ubiquitin ligase CUL7 revealed that MRN subunits are stable in cells infected with mutant virus or transfected with mutant viral DNA, implicating CUL7 association with T antigen in MRN proteolysis. The mutant genomes produce fewer virus progeny than the wild type, suggesting that T antigen-CUL7-directed proteolysis facilitates virus propagation. Use of a specific ATM kinase inhibitor showed that ATM kinase signaling is a prerequisite for proteasome-dependent degradation of MRN subunits as well as for the localization of T antigen and damage-signaling proteins to viral replication foci and optimal viral DNA replication. Taken together, the results indicate that SV40 infection manipulates host DNA damage-signaling to reprogram the cell for viral replication, perhaps through mechanisms related to host recovery from DNA damage.

UR - http://www.scopus.com/inward/record.url?scp=43949088506&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=43949088506&partnerID=8YFLogxK

U2 - 10.1128/JVI.02677-07

DO - 10.1128/JVI.02677-07

M3 - Article

C2 - 18353955

AN - SCOPUS:43949088506

VL - 82

SP - 5316

EP - 5328

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 11

ER -