Genotoxin-induced Rad9-Hus1-Rad1 (9-1-1) chromatin association is an early checkpoint signaling event

Pia Roos-Mattjus, Benjamin T. Vroman, Matthew A. Burtelow, Matthew Rauen, Alex K. Eapen, Larry M Karnitz

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Rad17, Rad1, Hus1, and Rad9 are key participants in checkpoint signaling pathways that block cell cycle progression in response to genotoxins. Biochemical and molecular modeling data predict that Rad9, Hus1, and Rad1 form a heterotrimeric complex, dubbed 9-1-1, which is loaded onto chromatin by a complex of Rad17 and the four small replication factor C (RFC) subunits (Rad17-RFC) in response to DNA damage. It is unclear what checkpoint proteins or checkpoint signaling events regulate the association of the 9-1-1 complex with DNA. Here we show that genotoxin-induced chromatin binding of 9-1-1 does not require the Rad9-inducible phosphorylation site (Ser-272). Although we found that Rad9 undergoes an additional phosphatidylinositol 3-kinase-related kinase (PIKK)-dependent posttranslational modification, we also show that genotoxin-triggered 9-1-1 chromatin binding does not depend on the catalytic activity of the PIKKs ataxia telangiectasia-mutated (ATM), ataxia telangiectasia and Rad3-related (ATR), or DNA-PK. Additionally, 9-1-1 chromatin binding does not require DNA replication, suggesting that the complex can be loaded onto DNA in response to DNA structures other than stalled DNA replication forks. Collectively, these studies demonstrate that 9-1-1 chromatin binding is a proximal event in the checkpoint signaling cascade.

Original languageEnglish (US)
Pages (from-to)43809-43812
Number of pages4
JournalJournal of Biological Chemistry
Volume277
Issue number46
DOIs
StatePublished - Nov 15 2002

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Mutagens
Chromatin
Association reactions
Replication Protein C
DNA
Ataxia Telangiectasia
DNA Replication
Phosphatidylinositol 3-Kinase
Post Translational Protein Processing
DNA Damage
Phosphorylation
Molecular modeling
Cell Cycle
Phosphotransferases
Catalyst activity
Cells
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Genotoxin-induced Rad9-Hus1-Rad1 (9-1-1) chromatin association is an early checkpoint signaling event. / Roos-Mattjus, Pia; Vroman, Benjamin T.; Burtelow, Matthew A.; Rauen, Matthew; Eapen, Alex K.; Karnitz, Larry M.

In: Journal of Biological Chemistry, Vol. 277, No. 46, 15.11.2002, p. 43809-43812.

Research output: Contribution to journalArticle

Roos-Mattjus, Pia ; Vroman, Benjamin T. ; Burtelow, Matthew A. ; Rauen, Matthew ; Eapen, Alex K. ; Karnitz, Larry M. / Genotoxin-induced Rad9-Hus1-Rad1 (9-1-1) chromatin association is an early checkpoint signaling event. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 46. pp. 43809-43812.
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