Mutational Analysis of the Herpes Simplex Virus Type 1 DNA Packaging Protein UL33
Mutational Analysis of the Herpes Simplex Virus Type 1 DNA Packaging Protein UL33
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DOI：
10.1128/jvi.01048-09
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发表时间：
2009-09-01
影响因子：
5.4
通讯作者：
Stow, Nigel D.
中科院分区：
文献类型：
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作者：
Beilstein, Frauke;Higgs, Martin R.;Stow, Nigel D.
单纯疱疹病毒1型（HSV-1）的UL33蛋白被认为是介导病毒DNA切割和包装的末端酶复合体的一个组成部分。在这项研究中，我们描述了一系列15个UL33突变体的产生和特征，这些突变体含有在130个残基蛋白中随机分布的5个氨基酸插入。在这些突变体中，有7个突变体在瞬时试验中不能补充ul33无效病毒的生长，也不能支持复制的扩增子DNA的切割和包装到衣壳中。这些突变体中的插入集中在51和74残基之间以及104和116残基之间，位于蛋白质最高度保守的区域。突变体与末端酶UL28组分相互作用的能力通过免疫沉淀和免疫荧光测定来评估。在氨基酸51和74之间插入的所有四个突变体在这种相互作用中都受到损害，而第二个区域的三个突变体中的两个（在位置111和116插入）不受影响。这些数据表明，UL33与UL28相互作用的能力可能是必要的，但不是充分的，以支持病毒生长和DNA包装。
The UL33 protein of herpes simplex virus type 1 (HSV-1) is thought to be a component of the terminase complex that mediates the cleavage and packaging of viral DNA. In this study we describe the generation and characterization of a series of 15 UL33 mutants containing insertions of five amino acids located randomly throughout the 130-residue protein. Of these mutants, seven were unable to complement the growth of the UL33-null virus dlUL33 in transient assays and also failed to support the cleavage and packaging of replicated amplicon DNA into capsids. The insertions in these mutants were clustered between residues 51 and 74 and between 104 and 116, within the most highly conserved regions of the protein. The ability of the mutants to interact with the UL28 component of the terminase was assessed in immunoprecipitation and immunofluorescence assays. All four mutants with insertions between amino acids 51 and 74 were impaired in this interaction, whereas two of the three mutants in the second region ( with insertions at positions 111 and 116) were not affected. These data indicate that the ability of UL33 to interact with UL28 is probably necessary, but not sufficient, to support viral growth and DNA packaging.