p48 activates a UV-damaged-DNA binding factor and is defective in xeroderma pigmentosum group E cells that lack binding activity

被引:139
作者
Hwang, BJ
Toering, S
Francke, U
Chu, G
机构
[1] Stanford Univ, Med Ctr,Sch Med, Dept Med, Div Oncol, Stanford, CA 94305 USA
[2] Stanford Univ, Med Ctr, Sch Med, Dept Biochem, Stanford, CA 94305 USA
[3] Stanford Univ, Med Ctr, Sch Med, Howard Hughes Med Inst, Stanford, CA 94305 USA
[4] Stanford Univ, Med Ctr, Sch Med, Dept Genet, Stanford, CA 94305 USA
关键词
D O I
10.1128/MCB.18.7.4391
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
A subset of xeroderma pigmentosum (XP) group E cells lack a factor that binds to DNA damaged by UV radiation. This factor can be purified to homogeneity as p125, a 125-kDa polypeptide, However, when cDNA encoding p125 is translated in vitro, only a small fraction binds to UV-damaged DNA, suggesting that a second factor is required for the activation of p125, We discovered that most hamster cell lines expressed inactive p125, which was activated in somatic cell hybrids containing human chromosome region 11p11.2-11cen. This region excluded p125 but included p48, which encodes a 48-kDa polypeptide known to copurify with p125 under some conditions. Expression of human p48 activated p125 binding in hamster cells and increased p125 binding in human cells. No such effects were observed from expression of p48 containing single amino acid substitutions from XP group E cells that lacked binding activity, demonstrating that the p48 gene is defective in those cells. Activation of p125 occurred by a "hit-and-run" mechanism, since the presence of p48 was not required for subsequent binding. Nevertheless, p48 was capable of forming a complex with p125 either bound to UV-damaged DNA or in free solution. It is notable that hamster cells fail to efficiently repair cyclobutane pyrimidine dimers in nontranscribed DNA and fail to express p48, which contains a WD motif with homology to proteins that reorganize chromatin, We propose that p48 plays a role in repairing lesions that would otherwise remain inaccessible in nontranscribed chromatin.
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页码:4391 / 4399
页数:9
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