Preferential binding of yeast Rad4•Rad23 complex to damaged DNA

被引：73

作者：

Jansen, LET ^{[1
]}

Verhage, RA ^{[1
]}

Brouwer, JA ^{[1
]}

机构：

[1] Leiden Univ, Leiden Inst Chem, MGC Dept Mol Genet, NL-2300 RA Leiden, Netherlands

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 1998年 / 273卷 / 50期

关键词：

D O I：

10.1074/jbc.273.50.33111

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The yeast Rad4 and Rad23 proteins form a complex that is involved in nucleotide excision repair (NER). Their function in this process is not known yet, but genetic data suggest that they act in an early step in NER. We have purified an epitope-tagged Rad4.Rad23 (tRad4.Rad23) complex from yeast cells, using a clone overproducing Rad4 with a hemagglutinin-tag at its C terminus. tRad4.Rad23 complex purified by both conventional and immuno-affinity chromatography complements the in vitro repair defect of rad4 and rad23 mutant extracts, demonstrating that these proteins are functional in NER. Using electrophoretic mobility shift assays, we show preferential binding of the tRad4.Rad23 complex to damaged DNA in vitro. UV-irradiated, as well as N-acetoxy-2-(acetylamino)fluorene-treated DNA, is efficiently bound by the protein complex. These data suggest that Rad4.Rad23 interacts with DNA damage during NER and may play a role in recognition of the damage.

引用

页码：33111 / 33114

页数：4

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