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PMID: 16713580 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

ATR kinase activation mediated by MutSalpha and MutLalpha in response to cytotoxic O6-methylguanine adducts.

Molecular cell ·Vol. 22 ·No. 4 ·2006-05-19 ·Pages 501-10

Yoshioka K, Yoshioka Y, Hsieh P

Abstract

S(N)1-type alkylating agents that produce cytotoxic O(6)-methyl-G (O(6)-meG) DNA adducts induce cell cycle arrest and apoptosis in a manner requiring the DNA mismatch repair (MMR) proteins MutSalpha and MutLalpha. Here, we show that checkpoint signaling in response to DNA methylation occurs during S phase and requires DNA replication that gives rise to O(6)-meG/T mispairs. DNA binding studies reveal that MutSalpha specifically recognizes O(6)-meG/T mispairs, but not O(6)-meG/C. In an in vitro assay, ATR-ATRIP, but not RPA, is preferentially recruited to O(6)-meG/T mismatches in a MutSalpha- and MutLalpha-dependent manner. Furthermore, ATR kinase is activated to phosphorylate Chk1 in the presence of O(6)-meG/T mispairs and MMR proteins. These results suggest that MMR proteins can act as direct sensors of methylation damage and help recruit ATR-ATRIP to sites of cytotoxic O(6)-meG adducts to initiate ATR checkpoint signaling.

MeSH Terms
Adaptor Proteins, Signal Transducing Animals Ataxia Telangiectasia Mutated Proteins Base Pair Mismatch Base Sequence Carrier Proteins/metabolism Cell Cycle Proteins/metabolism Cell Line DNA/genetics,metabolism DNA Adducts/metabolism,toxicity DNA Damage DNA Repair/physiology DNA-Binding Proteins Enzyme Activation/drug effects Exodeoxyribonucleases/metabolism Guanine/analogs & derivatives,metabolism HeLa Cells Humans Mice MutS DNA Mismatch-Binding Protein/metabolism Nuclear Proteins/metabolism Phosphoproteins/metabolism Protein Serine-Threonine Kinases/metabolism S Phase Signal Transduction
Chemicals
ATRIP protein, human Adaptor Proteins, Signal Transducing Carrier Proteins Cell Cycle Proteins DNA Adducts DNA-Binding Proteins Nuclear Proteins Phosphoproteins Guanine DNA O-(6)-methylguanine ATR protein, human Ataxia Telangiectasia Mutated Proteins Protein Serine-Threonine Kinases Exodeoxyribonucleases three prime repair exonuclease 1 MutS DNA Mismatch-Binding Protein
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Yoshioka Ken-ichi
Genetics and Biochemistry Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Yoshioka Yoshiko
Hsieh Peggy
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Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-2765
DOI
10.1016/j.molcel.2006.04.023
Published
2006-05-19
Pages
501-10
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC2423943
Subset
IM
Grants
Intramural NIH HHS · Z01 DK052029-01 · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2006.04.023 PMC Full Text View journal details
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