Gao M, DiTusa S F, Cordingley M G
Department of Virology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543.
Virology. 1993 Jun;194(2):647-53. doi: 10.1006/viro.1993.1304.
UL42 is the herpes simplex virus type 1 DNA polymerase (Pol) accessory protein and is required for viral DNA replication and growth. Previous results from this laboratory demonstrated that the N-terminal two thirds of the protein contains all of the biochemical activities of the protein which can be measured in vitro. These activities include dsDNA-binding, association with DNA polymerase, and stimulation of polymerase activity. To better understand the functions of UL42 in infected cells, we have isolated and characterized two viral recombinants, UL42lacZ and n338. In the mutant virus UL42lacZ, the UL42 gene was disrupted by insertion of the Escherichia coli lacZ gene, while in the mutant virus n338, a termination codon was introduced after amino acid position 338. Analysis of the mutant phenotypes suggest that (1) the first 338 residues of UL42 retain all the functions necessary for viral DNA replication and growth in lytic infection, (2) localization of UL42 to the cell nucleus is independent of Pol, and (3) localization of ICP8 (ssDNA-binding protein) to prereplication sites is independent of functional UL42.
UL42是单纯疱疹病毒1型DNA聚合酶(Pol)的辅助蛋白,是病毒DNA复制和生长所必需的。该实验室先前的结果表明,该蛋白的N端三分之二包含了可在体外测量的该蛋白的所有生化活性。这些活性包括双链DNA结合、与DNA聚合酶的结合以及对聚合酶活性的刺激。为了更好地理解UL42在受感染细胞中的功能,我们分离并鉴定了两种病毒重组体,即UL42lacZ和n338。在突变病毒UL42lacZ中,UL42基因因插入大肠杆菌lacZ基因而被破坏,而在突变病毒n338中,在氨基酸位置338之后引入了终止密码子。对突变表型的分析表明:(1)UL42的前338个残基保留了病毒DNA复制和裂解感染中生长所需的所有功能;(2)UL42定位于细胞核与Pol无关;(3)ICP8(单链DNA结合蛋白)定位于复制前位点与功能性UL42无关。