ESCHERICHIA-COLI UVRB PROTEIN BINDS TO DNA IN THE PRESENCE OF UVRA PROTEIN
ESCHERICHIA-COLI UVRB PROTEIN BINDS TO DNA IN THE PRESENCE OF UVRA PROTEIN
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DOI：
10.1038/294480a0
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发表时间：
1981-01-01
期刊：
影响因子：
64.8
通讯作者：
RUPP, WD
中科院分区：
文献类型：
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作者：
KACINSKI, BM;RUPP, WD
在体内和体外，含有嘧啶二聚体或其他大块病变的受损DNA的核酸内切酶切割需要大肠杆菌的uvrA、uvrBanduvrC基因产物的作用1 -3。为了更好地理解这三个基因的产物如何介导切口，已经克隆了所有三个基因4 -10。虽然这三个基因（命名为UVRA、UVRB和UVRC）的基因产物在核苷酸切除修复的早期阶段都是必需的，但尚不清楚它们是独立发挥作用还是作为多蛋白复合物的一部分一起发挥作用。由于这三种蛋白质参与受损DNA的修复，因此有理由预期在某些情况下所有三种蛋白质都应与DNA结合，并且与此预期雅阁，先前的实验已经证明UVRA和UVRC与单链DNA紧密结合11 -13。我们已经调查了是否UVRB也是一种DNA结合蛋白，并在这里报告的UVRB的放射化学纯化和实验使用这种放射性标记的UVRB来测量其结合到DNA。UVRB本身不与DNA强烈结合，但在UVRA存在下，观察到UVRB与单链DNA的紧密结合，从而证明UVRA和UVRB之间发生直接相互作用。
In vivoandin vitro, the endonucleolytic incision of damaged DNA containing pyrimidine dimers or other bulky lesions requires the action of theuvrA,uvrBanduvrCgene products ofEscherichia coli1–3. To understand better how the products of these three genes mediate incision, all three genes have been cloned4–10. Although the gene products of the three genes (designated UVRA, UVRB and UVRC) are all required at the early stages of nucleotide excision repair, it is not known whether they act independently or together as part of a multi-protein complex. As the three proteins are involved in the repair of damaged DNA, it is reasonable to expect that under some circumstances all three proteins should bind to DNA, and, in accord with this expectation, previous experiments have demonstrated that UVRA and UVRC bind tightly to single-stranded DNA11–13. We have investigated whether UVRB is also a DNA-binding protein and report here a radiochemical purification of UVRB and experiments using this radiolabelled UVRB to measure its binding to DNA. By itself, UVRB does not bind strongly to DNA, but in the presence of UVRA, tight binding of UVRB to single-stranded DNA is observed, thus demonstrating the occurrence of direct interactions between UVRA and UVRB.