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Nucleic Acids Research, 2000, Vol. 28, No. 23 4800-4804
© 2000 Oxford University Press

Differential processing of UV mimetic and interstrand crosslink damage by XPF cell extracts

Nianxiang Zhang, Xiaoshan Zhang, Carolyn Peterson, Lei Li1 and Randy Legerski*

Department of Molecular Genetics and 1Department of Experimental Radiation Oncology, The University of Texas, M.D. Anderson Cancer Center, Houston, TX 77030, USA

We have recently developed a mammalian cell free assay in which interstrand crosslinks induce DNA synthesis in both damaged and undamaged plasmids co-incubated in the same extract. We have also shown using hamster mutants that both ERCC1 and XPF are required for the observed incorporation. Here, we show that extracts from an XPF patient cell line differentially process UV mimetic damage and interstrand crosslinks in vitro. XPF extracts are highly defective in the stimulation of repair synthesis by N-acetoxy-N- acetylaminofluorene, but are proficient in the stimulation of DNA synthesis by psoralen interstrand crosslinks. In addition, we show that extracts from the hamster UV140 mutant, which has high UV sensitivity, but moderate mitomycin C sensitivity, are similar in both assays to XPF cell extracts. These findings support the hypothesis that the activities of XPF in nucleotide excision repair (NER) and crosslink repair are separable, and that mutations in XPF patients result in the abolition of NER, but not recombinational repair pathways, which are likely to be essential as has been observed in ERCC1 homozygous –/– mice.

* To whom correspondence should be addressed. Tel: +1 713 792 8941; Fax: +1 713 794 4295; Email: rlegersk{at}mdanderson.org


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