Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway

Junya Kobayashi, Hiroko Fujimoto, Jun Sato, Ikue Hayashi, Sandeep Burma, Shinya Matsuura, David J. Chen, Kenshi Komatsu

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

H2AX is an important factor for chromatin remodeling to facilitate accumulation of DNA damage-related proteins at DNA double-strand break (DSB) sites. In order to further understand the role of H2AX in the DNA damage response (DDR), we attempted to identify H2AX-interacting proteins by proteomics analysis. As a result, we identified nucleolin as one of candidates. Here, we show a novel role of a major nucleolar protein, nucleolin, in DDR. Nucleolin interacted with γ-H2AX and accumulated to laser micro-irradiated DSB damage sites. Chromatin Immunoprecipitation assay also displayed the accumulation of nucleolin around DSB sites. Nucleolin-depleted cells exhibited repression of both ATM-dependent phosphorylation following exposure to γ-ray and subsequent cell cycle checkpoint activation. Furthermore, nucleolin-knockdown reduced HR and NHEJ activity and showed decrease in IR-induced chromatin accumulation of HR/NHEJ factors, agreeing with the delayed kinetics of γ-H2AX focus. Moreover, nucleolin-knockdown decreased MDC1-related events such as focus formation of 53 BP1, RNF168, phosphorylated ATM, and H2A ubiquitination. Nucleolin also showed FACT-like activity for DSB damage-induced histone eviction from chromatin. Taken together, nucleolin could promote both ATM-dependent cell cycle checkpoint and DSB repair by functioning in an MDC1-related pathway through its FACT-like function.

Original languageEnglish (US)
Article numbere49245
JournalPLoS One
Volume7
Issue number11
DOIs
StatePublished - Nov 7 2012

Fingerprint

Double-Stranded DNA Breaks
chromatin
DNA damage
DNA
Chromatin
Automatic teller machines
DNA Damage
proteins
histones
proteomics
Cell Cycle Checkpoints
lasers
phosphorylation
Cells
kinetics
nucleolin
Phosphorylation
assays
Chromatin Assembly and Disassembly
Chromatin Immunoprecipitation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Kobayashi, J., Fujimoto, H., Sato, J., Hayashi, I., Burma, S., Matsuura, S., ... Komatsu, K. (2012). Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway. PLoS One, 7(11), [e49245]. https://doi.org/10.1371/journal.pone.0049245

Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway. / Kobayashi, Junya; Fujimoto, Hiroko; Sato, Jun; Hayashi, Ikue; Burma, Sandeep; Matsuura, Shinya; Chen, David J.; Komatsu, Kenshi.

In: PLoS One, Vol. 7, No. 11, e49245, 07.11.2012.

Research output: Contribution to journalArticle

Kobayashi, J, Fujimoto, H, Sato, J, Hayashi, I, Burma, S, Matsuura, S, Chen, DJ & Komatsu, K 2012, 'Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway', PLoS One, vol. 7, no. 11, e49245. https://doi.org/10.1371/journal.pone.0049245
Kobayashi, Junya ; Fujimoto, Hiroko ; Sato, Jun ; Hayashi, Ikue ; Burma, Sandeep ; Matsuura, Shinya ; Chen, David J. ; Komatsu, Kenshi. / Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway. In: PLoS One. 2012 ; Vol. 7, No. 11.
@article{7e75d2564c3349339063db4136316be2,
title = "Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway",
abstract = "H2AX is an important factor for chromatin remodeling to facilitate accumulation of DNA damage-related proteins at DNA double-strand break (DSB) sites. In order to further understand the role of H2AX in the DNA damage response (DDR), we attempted to identify H2AX-interacting proteins by proteomics analysis. As a result, we identified nucleolin as one of candidates. Here, we show a novel role of a major nucleolar protein, nucleolin, in DDR. Nucleolin interacted with γ-H2AX and accumulated to laser micro-irradiated DSB damage sites. Chromatin Immunoprecipitation assay also displayed the accumulation of nucleolin around DSB sites. Nucleolin-depleted cells exhibited repression of both ATM-dependent phosphorylation following exposure to γ-ray and subsequent cell cycle checkpoint activation. Furthermore, nucleolin-knockdown reduced HR and NHEJ activity and showed decrease in IR-induced chromatin accumulation of HR/NHEJ factors, agreeing with the delayed kinetics of γ-H2AX focus. Moreover, nucleolin-knockdown decreased MDC1-related events such as focus formation of 53 BP1, RNF168, phosphorylated ATM, and H2A ubiquitination. Nucleolin also showed FACT-like activity for DSB damage-induced histone eviction from chromatin. Taken together, nucleolin could promote both ATM-dependent cell cycle checkpoint and DSB repair by functioning in an MDC1-related pathway through its FACT-like function.",
author = "Junya Kobayashi and Hiroko Fujimoto and Jun Sato and Ikue Hayashi and Sandeep Burma and Shinya Matsuura and Chen, {David J.} and Kenshi Komatsu",
year = "2012",
month = "11",
day = "7",
doi = "10.1371/journal.pone.0049245",
language = "English (US)",
volume = "7",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "11",

}

TY - JOUR

T1 - Nucleolin Participates in DNA Double-Strand Break-Induced Damage Response through MDC1-Dependent Pathway

AU - Kobayashi, Junya

AU - Fujimoto, Hiroko

AU - Sato, Jun

AU - Hayashi, Ikue

AU - Burma, Sandeep

AU - Matsuura, Shinya

AU - Chen, David J.

AU - Komatsu, Kenshi

PY - 2012/11/7

Y1 - 2012/11/7

N2 - H2AX is an important factor for chromatin remodeling to facilitate accumulation of DNA damage-related proteins at DNA double-strand break (DSB) sites. In order to further understand the role of H2AX in the DNA damage response (DDR), we attempted to identify H2AX-interacting proteins by proteomics analysis. As a result, we identified nucleolin as one of candidates. Here, we show a novel role of a major nucleolar protein, nucleolin, in DDR. Nucleolin interacted with γ-H2AX and accumulated to laser micro-irradiated DSB damage sites. Chromatin Immunoprecipitation assay also displayed the accumulation of nucleolin around DSB sites. Nucleolin-depleted cells exhibited repression of both ATM-dependent phosphorylation following exposure to γ-ray and subsequent cell cycle checkpoint activation. Furthermore, nucleolin-knockdown reduced HR and NHEJ activity and showed decrease in IR-induced chromatin accumulation of HR/NHEJ factors, agreeing with the delayed kinetics of γ-H2AX focus. Moreover, nucleolin-knockdown decreased MDC1-related events such as focus formation of 53 BP1, RNF168, phosphorylated ATM, and H2A ubiquitination. Nucleolin also showed FACT-like activity for DSB damage-induced histone eviction from chromatin. Taken together, nucleolin could promote both ATM-dependent cell cycle checkpoint and DSB repair by functioning in an MDC1-related pathway through its FACT-like function.

AB - H2AX is an important factor for chromatin remodeling to facilitate accumulation of DNA damage-related proteins at DNA double-strand break (DSB) sites. In order to further understand the role of H2AX in the DNA damage response (DDR), we attempted to identify H2AX-interacting proteins by proteomics analysis. As a result, we identified nucleolin as one of candidates. Here, we show a novel role of a major nucleolar protein, nucleolin, in DDR. Nucleolin interacted with γ-H2AX and accumulated to laser micro-irradiated DSB damage sites. Chromatin Immunoprecipitation assay also displayed the accumulation of nucleolin around DSB sites. Nucleolin-depleted cells exhibited repression of both ATM-dependent phosphorylation following exposure to γ-ray and subsequent cell cycle checkpoint activation. Furthermore, nucleolin-knockdown reduced HR and NHEJ activity and showed decrease in IR-induced chromatin accumulation of HR/NHEJ factors, agreeing with the delayed kinetics of γ-H2AX focus. Moreover, nucleolin-knockdown decreased MDC1-related events such as focus formation of 53 BP1, RNF168, phosphorylated ATM, and H2A ubiquitination. Nucleolin also showed FACT-like activity for DSB damage-induced histone eviction from chromatin. Taken together, nucleolin could promote both ATM-dependent cell cycle checkpoint and DSB repair by functioning in an MDC1-related pathway through its FACT-like function.

UR - http://www.scopus.com/inward/record.url?scp=84868681561&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84868681561&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0049245

DO - 10.1371/journal.pone.0049245

M3 - Article

C2 - 23145133

AN - SCOPUS:84868681561

VL - 7

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 11

M1 - e49245

ER -