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Molecular and Cellular Biology, January 2003, p. 754-761, Vol. 23, No. 2
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.2.754-761.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Nucleotide Excision Repair- and Polymerase {eta}-Mediated Error-Prone Removal of Mitomycin C Interstrand Cross-Links

Huyong Zheng,1 Xin Wang,1 Amy J. Warren,2 Randy J. Legerski,3 Rodney S. Nairn,4 Joshua W. Hamilton,2 and Lei Li1,3*

Departments of Experimental Radiation Oncology,1 Molecular Genetics,3 Carcinogenesis, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030,4 Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 037552

Received 28 December 2001/ Returned for modification 4 February 2002/ Accepted 10 October 2002

Interstrand cross-links (ICLs) make up a unique class of DNA lesions in which both strands of the double helix are covalently joined, precluding strand opening during replication and transcription. The repair of DNA ICLs has become a focus of study since ICLs are recognized as the main cytotoxic lesion inflicted by an array of alkylating compounds used in cancer treatment. As is the case for double-strand breaks, a damage-free homologous copy is essential for the removal of ICLs in an error-free manner. However, recombination-independent mechanisms may exist to remove ICLs in an error-prone fashion. We have developed an in vivo reactivation assay that can be used to examine the removal of site-specific mitomycin C-mediated ICLs in mammalian cells. We found that the removal of the ICL from the reporter substrate could take place in the absence of undamaged homologous sequences in repair-proficient cells, suggesting a cross-link repair mechanism that is independent of homologous recombination. Systematic analysis of nucleotide excision repair mutants demonstrated the involvement of transcription-coupled nucleotide excision repair and a partial requirement for the lesion bypass DNA polymerase {eta} encoded by the human POLH gene. From these observations, we propose the existence of a recombination-independent and mutagenic repair pathway for the removal of ICLs in mammalian cells.

* Corresponding author. Mailing address: Department of Experimental Radiation Oncology, The University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX 77030. Phone: (713) 792-3424. Fax: (716) 794-5369. E-mail: leili{at}mdanderson.org.

Molecular and Cellular Biology, January 2003, p. 754-761, Vol. 23, No. 2
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.2.754-761.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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