Identification of a coiled coil in Werner syndrome protein that facilitates multimerization and promotes exonuclease processivity

J. Jefferson P Perry, Aroumougame Asaithamby, Adam Barnebey, Foad Kiamanesch, David J. Chen, Seungil Han, John A. Tainer, Steven M. Yannone

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Werner syndrome (WS) is a rare progeroid disorder characterized by genomic instability, increased cancer incidence, and early onset of a variety of aging pathologies. WS is unique among early aging syndromes in that affected individuals are developmentally normal, and phenotypic onset is in early adulthood. The protein defective in WS (WRN) is a member of the large RecQ family of helicases but is unique among this family in having an exonuclease. RecQ helicases form multimers, but the mechanism and consequence of multimerization remain incompletely defined. Here, we identify a novel heptad repeat coiled coil region between the WRN nuclease and helicase domains that facilitates multimerization of WRN. We mapped a novel and unique DNA-dependent protein kinase phosphorylation site proximal to the WRN multimerization region. However, phosphorylation at this site affected neither exonuclease activity nor multimericstate. We found that WRN nuclease is stimulated by DNA-dependent protein kinase independently of kinase activity or WRN nuclease multimeric status. In addition, WRN nuclease multimerization significantly increased nuclease processivity. We found that the novel WRN coiled coil domain is necessary for multimerization of the nuclease domain and sufficient to multimerize with full-length WRN in human cells. Importantly, correct homomultimerization is required for WRN function in vivo as overexpression of this multimerization domain caused increased sensitivity to camptothecin and 4-nitroquinoline 1-oxide similar to that in cells lacking functional WRN protein.

Original languageEnglish (US)
Pages (from-to)25699-25707
Number of pages9
JournalJournal of Biological Chemistry
Volume285
Issue number33
DOIs
StatePublished - Aug 13 2010

Fingerprint

Protein Multimerization
DNA-Activated Protein Kinase
Werner Syndrome
Exonucleases
Phosphorylation
RecQ Helicases
Aging of materials
4-Nitroquinoline-1-oxide
Camptothecin
Pathology
Proteins
Phosphotransferases
Genomic Instability
Cells
Incidence
Neoplasms

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Identification of a coiled coil in Werner syndrome protein that facilitates multimerization and promotes exonuclease processivity. / Perry, J. Jefferson P; Asaithamby, Aroumougame; Barnebey, Adam; Kiamanesch, Foad; Chen, David J.; Han, Seungil; Tainer, John A.; Yannone, Steven M.

In: Journal of Biological Chemistry, Vol. 285, No. 33, 13.08.2010, p. 25699-25707.

Research output: Contribution to journalArticle

Perry, J. Jefferson P ; Asaithamby, Aroumougame ; Barnebey, Adam ; Kiamanesch, Foad ; Chen, David J. ; Han, Seungil ; Tainer, John A. ; Yannone, Steven M. / Identification of a coiled coil in Werner syndrome protein that facilitates multimerization and promotes exonuclease processivity. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 33. pp. 25699-25707.
@article{d15264012fd04267b3031c7c05aa00f2,
title = "Identification of a coiled coil in Werner syndrome protein that facilitates multimerization and promotes exonuclease processivity",
abstract = "Werner syndrome (WS) is a rare progeroid disorder characterized by genomic instability, increased cancer incidence, and early onset of a variety of aging pathologies. WS is unique among early aging syndromes in that affected individuals are developmentally normal, and phenotypic onset is in early adulthood. The protein defective in WS (WRN) is a member of the large RecQ family of helicases but is unique among this family in having an exonuclease. RecQ helicases form multimers, but the mechanism and consequence of multimerization remain incompletely defined. Here, we identify a novel heptad repeat coiled coil region between the WRN nuclease and helicase domains that facilitates multimerization of WRN. We mapped a novel and unique DNA-dependent protein kinase phosphorylation site proximal to the WRN multimerization region. However, phosphorylation at this site affected neither exonuclease activity nor multimericstate. We found that WRN nuclease is stimulated by DNA-dependent protein kinase independently of kinase activity or WRN nuclease multimeric status. In addition, WRN nuclease multimerization significantly increased nuclease processivity. We found that the novel WRN coiled coil domain is necessary for multimerization of the nuclease domain and sufficient to multimerize with full-length WRN in human cells. Importantly, correct homomultimerization is required for WRN function in vivo as overexpression of this multimerization domain caused increased sensitivity to camptothecin and 4-nitroquinoline 1-oxide similar to that in cells lacking functional WRN protein.",
author = "Perry, {J. Jefferson P} and Aroumougame Asaithamby and Adam Barnebey and Foad Kiamanesch and Chen, {David J.} and Seungil Han and Tainer, {John A.} and Yannone, {Steven M.}",
year = "2010",
month = "8",
day = "13",
doi = "10.1074/jbc.M110.124941",
language = "English (US)",
volume = "285",
pages = "25699--25707",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "33",

}

TY - JOUR

T1 - Identification of a coiled coil in Werner syndrome protein that facilitates multimerization and promotes exonuclease processivity

AU - Perry, J. Jefferson P

AU - Asaithamby, Aroumougame

AU - Barnebey, Adam

AU - Kiamanesch, Foad

AU - Chen, David J.

AU - Han, Seungil

AU - Tainer, John A.

AU - Yannone, Steven M.

PY - 2010/8/13

Y1 - 2010/8/13

N2 - Werner syndrome (WS) is a rare progeroid disorder characterized by genomic instability, increased cancer incidence, and early onset of a variety of aging pathologies. WS is unique among early aging syndromes in that affected individuals are developmentally normal, and phenotypic onset is in early adulthood. The protein defective in WS (WRN) is a member of the large RecQ family of helicases but is unique among this family in having an exonuclease. RecQ helicases form multimers, but the mechanism and consequence of multimerization remain incompletely defined. Here, we identify a novel heptad repeat coiled coil region between the WRN nuclease and helicase domains that facilitates multimerization of WRN. We mapped a novel and unique DNA-dependent protein kinase phosphorylation site proximal to the WRN multimerization region. However, phosphorylation at this site affected neither exonuclease activity nor multimericstate. We found that WRN nuclease is stimulated by DNA-dependent protein kinase independently of kinase activity or WRN nuclease multimeric status. In addition, WRN nuclease multimerization significantly increased nuclease processivity. We found that the novel WRN coiled coil domain is necessary for multimerization of the nuclease domain and sufficient to multimerize with full-length WRN in human cells. Importantly, correct homomultimerization is required for WRN function in vivo as overexpression of this multimerization domain caused increased sensitivity to camptothecin and 4-nitroquinoline 1-oxide similar to that in cells lacking functional WRN protein.

AB - Werner syndrome (WS) is a rare progeroid disorder characterized by genomic instability, increased cancer incidence, and early onset of a variety of aging pathologies. WS is unique among early aging syndromes in that affected individuals are developmentally normal, and phenotypic onset is in early adulthood. The protein defective in WS (WRN) is a member of the large RecQ family of helicases but is unique among this family in having an exonuclease. RecQ helicases form multimers, but the mechanism and consequence of multimerization remain incompletely defined. Here, we identify a novel heptad repeat coiled coil region between the WRN nuclease and helicase domains that facilitates multimerization of WRN. We mapped a novel and unique DNA-dependent protein kinase phosphorylation site proximal to the WRN multimerization region. However, phosphorylation at this site affected neither exonuclease activity nor multimericstate. We found that WRN nuclease is stimulated by DNA-dependent protein kinase independently of kinase activity or WRN nuclease multimeric status. In addition, WRN nuclease multimerization significantly increased nuclease processivity. We found that the novel WRN coiled coil domain is necessary for multimerization of the nuclease domain and sufficient to multimerize with full-length WRN in human cells. Importantly, correct homomultimerization is required for WRN function in vivo as overexpression of this multimerization domain caused increased sensitivity to camptothecin and 4-nitroquinoline 1-oxide similar to that in cells lacking functional WRN protein.

UR - http://www.scopus.com/inward/record.url?scp=77955480303&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955480303&partnerID=8YFLogxK

U2 - 10.1074/jbc.M110.124941

DO - 10.1074/jbc.M110.124941

M3 - Article

VL - 285

SP - 25699

EP - 25707

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 33

ER -