Use of transdominant mutants of the origin-binding protein (UL9) of herpes simplex virus type 1 to define functional domains
Use of transdominant mutants of the origin-binding protein (UL9) of herpes simplex virus type 1 to define functional domains
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DOI：
10.1128/jvi.70.11.7859-7866.1996
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发表时间：
1996-11-01
影响因子：
5.4
通讯作者：
Weller, SK
中科院分区：
文献类型：
--
作者：
Malik, AK;Weller, SK
UL9是单纯疱疹病毒1型的起源结合蛋白，包含在RNA和DNA解旋酶的大超家族中保守的六个序列基序。在这些蛾中的单氨基酸取代突变在体内起作用（R Martinez，L. Shao和S. R Weller，J. Virol. 66：6735 - 6746，1992）。野生型UL9的过表达在转染测定中抑制空斑形成，所述转染测定测量感染性单纯疱疹病毒1型DNA的病毒空斑形成。在基序I、II或VI中含有突变的构建体在相同的测定中表现出甚至更强的抑制作用，因此可以认为是野生型病毒噬斑形成的强反显性抑制剂。可以通过在DNA结合结构域中引入第二突变或通过缺失蛋白质的N-末端35个氨基酸来减轻转显性表型。野生型UL9的抑制作用也可以通过缺失氨基酸292至404而部分减轻。我们提出UL9的N-末端35个氨基酸和残基292至404可能定义UL9蛋白的新功能结构域。
UL9, the origin-binding protein of herpes simplex virus type 1, contains six sequence motifs conserved in a large superfamily of RNA and DNA helicases. Single-amino-acid substitution mutations in these moths inactivate UL9 function in vivo (R Martinez, L. Shao, and S. R Weller, J. Virol. 66:6735-6746, 1992). Overexpression of wild-type UL9 is inhibitory to plaque formation in a transfection assay which measures viral plaque formation by infectious herpes simplex: virus type 1 DNA. Constructs containing mutations in motif I, II, or VI exhibit even stronger inhibitory effects in the same assay and thus can be considered strong transdominant inhibitors of plaque formation by the wild-type virus. The transdominant phenotype can be relieved by introducing a second mutation in the DNA-binding domain or by deleting the N-terminal 35 amino acids of the protein. The inhibitory effects of wild-type UL9 can also be partially relieved by deletion of amino acids 292 to 404. We propose that the N-terminal 35 amino acids of UL9 and residues 292 to 404 may define new functional domains of the UL9 protein.