Werner Syndrome Protein Is Regulated and Phosphorylated by DNA-dependent Protein Kinase

Steven M. Yannone, Sashwati Roy, Doug W. Chan, Michael B. Murphy, Shurong Huang, Judith Campisi, David J. Chen

Research output: Contribution to journalArticle

171 Citations (Scopus)

Abstract

DNA double-strand breaks (DSBs) are a highly mutagenic and potentially lethal damage that occurs in all organisms. Mammalian cells repair DSBs by homologous recombination and non-homologous end joining, the latter requiring DNA-dependent protein kinase (DNA-PK). Werner syndrome is a disorder characterized by genomic instability, aging pathologies and defective WRN, a RecQ-like helicase with exonuclease activity. We show that WRN interacts directly with the catalytic subunit of DNA-PK (DNA-PKCS), which inhibits both the helicase and exonuclease activities of WRN. In addition we show that WRN forms a stable complex on DNA with DNA-PKCS and the DNA binding subunit Ku. This assembly reverses WRN enzymatic inhibition. Finally, we show that WRN is phosphorylated in vitro by DNA-PK and requires DNA-PK for phosphorylation in vivo, and that cells deficient in WRN are mildly sensitive to ionizing radiation. These data suggest that DNA-PK and WRN may function together in DNA metabolism and implicate WRN function in non-homologous end joining.

Original languageEnglish (US)
Pages (from-to)38242-38248
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number41
StatePublished - Oct 12 2001

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DNA-Activated Protein Kinase
Werner Syndrome
lysyl-arginyl-alanyl-lysyl-alanyl-lysyl-threonyl-threonyl-lysyl-lysyl-arginine
DNA
Exonucleases
Proteins
Joining
RecQ Helicases
Catalytic DNA
Double-Stranded DNA Breaks
Genomic Instability
Homologous Recombination
Ionizing Radiation
Phosphorylation
Ionizing radiation
Pathology
Catalytic Domain
Metabolism
Repair
Aging of materials

ASJC Scopus subject areas

  • Biochemistry

Cite this

Yannone, S. M., Roy, S., Chan, D. W., Murphy, M. B., Huang, S., Campisi, J., & Chen, D. J. (2001). Werner Syndrome Protein Is Regulated and Phosphorylated by DNA-dependent Protein Kinase. Journal of Biological Chemistry, 276(41), 38242-38248.

Werner Syndrome Protein Is Regulated and Phosphorylated by DNA-dependent Protein Kinase. / Yannone, Steven M.; Roy, Sashwati; Chan, Doug W.; Murphy, Michael B.; Huang, Shurong; Campisi, Judith; Chen, David J.

In: Journal of Biological Chemistry, Vol. 276, No. 41, 12.10.2001, p. 38242-38248.

Research output: Contribution to journalArticle

Yannone, SM, Roy, S, Chan, DW, Murphy, MB, Huang, S, Campisi, J & Chen, DJ 2001, 'Werner Syndrome Protein Is Regulated and Phosphorylated by DNA-dependent Protein Kinase', Journal of Biological Chemistry, vol. 276, no. 41, pp. 38242-38248.
Yannone SM, Roy S, Chan DW, Murphy MB, Huang S, Campisi J et al. Werner Syndrome Protein Is Regulated and Phosphorylated by DNA-dependent Protein Kinase. Journal of Biological Chemistry. 2001 Oct 12;276(41):38242-38248.
Yannone, Steven M. ; Roy, Sashwati ; Chan, Doug W. ; Murphy, Michael B. ; Huang, Shurong ; Campisi, Judith ; Chen, David J. / Werner Syndrome Protein Is Regulated and Phosphorylated by DNA-dependent Protein Kinase. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 41. pp. 38242-38248.
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AB - DNA double-strand breaks (DSBs) are a highly mutagenic and potentially lethal damage that occurs in all organisms. Mammalian cells repair DSBs by homologous recombination and non-homologous end joining, the latter requiring DNA-dependent protein kinase (DNA-PK). Werner syndrome is a disorder characterized by genomic instability, aging pathologies and defective WRN, a RecQ-like helicase with exonuclease activity. We show that WRN interacts directly with the catalytic subunit of DNA-PK (DNA-PKCS), which inhibits both the helicase and exonuclease activities of WRN. In addition we show that WRN forms a stable complex on DNA with DNA-PKCS and the DNA binding subunit Ku. This assembly reverses WRN enzymatic inhibition. Finally, we show that WRN is phosphorylated in vitro by DNA-PK and requires DNA-PK for phosphorylation in vivo, and that cells deficient in WRN are mildly sensitive to ionizing radiation. These data suggest that DNA-PK and WRN may function together in DNA metabolism and implicate WRN function in non-homologous end joining.

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