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PMID: 14614141 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The base excision repair enzyme MED1 mediates DNA damage response to antitumor drugs and is associated with mismatch repair system integrity.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 100 ·No. 25 ·2003-12-09 ·Pages 15071-6

Cortellino S, Turner D, Masciullo V, Schepis F, Albino D, Daniel R, Skalka AM, Meropol NJ, Alberti C, Larue L, Bellacosa A

Abstract

Cytotoxicity of methylating agents is caused mostly by methylation of the O6 position of guanine in DNA to form O6-methylguanine (O6-meG). O6-meG can direct misincorporation of thymine during replication, generating O6-meG:T mismatches. Recognition of these mispairs by the mismatch repair (MMR) system leads to cell cycle arrest and apoptosis. MMR also modulates sensitivity to other antitumor drugs. The base excision repair (BER) enzyme MED1 (also known as MBD4) interacts with the MMR protein MLH1. MED1 was found to exhibit thymine glycosylase activity on O6-meG:T mismatches. To examine the biological significance of this activity, we generated mice with targeted inactivation of the Med1 gene and prepared mouse embryonic fibroblasts (MEF) with different Med1 genotype. Unlike wild-type and heterozygous cultures, Med1-/- MEF failed to undergo G2-M cell cycle arrest and apoptosis upon treatment with the methylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Similar results were obtained with platinum compounds' 5-fluorouracil and irinotecan. As is the case with MMR-defective cells, resistance of Med1-/- MEF to MNNG was due to a tolerance mechanism because DNA damage accumulated but did not elicit checkpoint activation. Interestingly, steady state amounts of several MMR proteins are reduced in Med1-/- MEF, in comparison with Med1+/+ and Med1+/- MEF. We conclude that MED1 has an additional role in DNA damage response to antitumor agents and is associated with integrity of the MMR system. MED1 defects (much like MMR defects) may impair cell cycle arrest and apoptosis induced by DNA damage.

MeSH Terms
Animals Antineoplastic Agents/pharmacology Apoptosis Base Pair Mismatch Blotting, Western Cell Separation Cells, Cultured DNA Damage DNA Methylation DNA Repair Endodeoxyribonucleases/genetics,physiology Fibroblasts/metabolism Flow Cytometry G2 Phase Genotype Guanine Methylnitronitrosoguanidine Mice Mice, Transgenic Mitosis Models, Genetic Retroviridae/genetics Reverse Transcriptase Polymerase Chain Reaction Thymine DNA Glycosylase/metabolism Time Factors
Chemicals
Antineoplastic Agents Methylnitronitrosoguanidine Guanine Endodeoxyribonucleases Mbd4 protein, mouse Thymine DNA Glycosylase
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Cortellino Salvatore
Program in Human Genetics, Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111, USA.
Turner David
Masciullo Valeria
Schepis Filippo
Albino Domenico
Daniel Rene
Skalka Anna Marie
Meropol Neal J
Alberti Christophe
Larue Lionel
Bellacosa Alfonso
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.2334585100
Published
2003-12-09
Epub
2003-00-12
Pages
15071-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC299910
Subset
IM
Grants
NCI NIH HHS · CA06927 · United States
NCI NIH HHS · R01 CA078412 · United States
NCI NIH HHS · R29 CA078412 · United States
NCI NIH HHS · CA78412 · United States
NCI NIH HHS · P30 CA006927 · United States
Corrections
CommentIn
PMID: 14657347
External Links
PubMed source DOI: 10.1073/pnas.2334585100 PMC Full Text View journal details
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