Photoaffinity labeling of the herpes simplex virus type-1 single-strand DNA-binding protein (ICP8) with oligodeoxyribonucleotides.
Photoaffinity labeling of the herpes simplex virus type-1 single-strand DNA-binding protein (ICP8) with oligodeoxyribonucleotides.
复制标题
用寡脱氧核糖核苷酸光亲和标记 1 型单纯疱疹病毒单链 DNA 结合蛋白 (ICP8)。
DOI：
10.1006/bbrc.1999.1566
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发表时间：
1999
影响因子：
3.1
通讯作者：
Boehmer,PE
中科院分区：
文献类型：
--
作者：
White,EJ;Boehmer,PE
单纯疱疹病毒1型单链DNA结合蛋白ICP 8是一种128 kDa的锌金属蛋白。在本通信中，我们已经表明，未取代的和溴脱氧尿苷取代的寡核苷酸可以通过UV照射特异性交联到ICP 8。我们已经使用这种方法来显示ICP 8的单链DNA结合位点位于53.5 kDa的胰蛋白酶多肽内。该多肽起始于丙氨酸368，估计延伸至精氨酸902。体外合成的包含残基368-902的多肽表现出单链DNA结合活性。我们的结论是，该区域包括残基368-902包含的单链DNA结合位点的ICP 8。此外，ICP 8与寡核苷酸的光亲和标记提供了一种特异性修饰其单链DNA结合位点的方法，从而促进了未来对其单链DNA结合活性在与其他DNA复制酶相互作用中的重要性的研究。
The herpes simplex virus type-1 single-strand DNA-binding protein ICP8 is a 128-kDa zinc metalloprotein. In this communication we have shown that unsubstituted and bromodeoxyuridine-substituted oligonucleotides can be specifically crosslinked to ICP8 by UV irradiation. We have used this approach to show that the single-strand DNA-binding site of ICP8 resides within a 53.5-kDa tryptic polypeptide. This polypeptide initiates at alanine 368 and was estimated to extend through arginine 902. A polypeptide encompassing residues 368–902 synthesized in vitro exhibited single-strand DNA-binding activity. We conclude that the region encompassing residues 368–902 contains the single-strand DNA-binding site of ICP8. Moreover, photoaffinity labeling of ICP8 with oligonucleotides provides a means of specifically modifying its single-strand DNA-binding site, thereby facilitating future studies on the importance of its single-strand DNA-binding activity in its interaction with other DNA replication enzymes.