TY - JOUR
T1 - Differential processing of UV mimetic and interstrand crosslink damage by XPF cell extracts
AU - Zhang, Nianxiang
AU - Zhang, Xiaoshan
AU - Peterson, Carolyn
AU - Li, Lei
AU - Legerski, Randy
PY - 2000/12/1
Y1 - 2000/12/1
N2 - 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 inter-strand 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.
AB - 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 inter-strand 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.
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U2 - 10.1093/nar/28.23.4800
DO - 10.1093/nar/28.23.4800
M3 - Article
C2 - 11095693
AN - SCOPUS:0034549346
SN - 0305-1048
VL - 28
SP - 4800
EP - 4804
JO - Nucleic acids research
JF - Nucleic acids research
IS - 23
ER -