The Herpes Simplex Virus Type 1 UL20 Protein and the Amino Terminus of Glycoprotein K (gK) Physically Interact with gB
The Herpes Simplex Virus Type 1 UL20 Protein and the Amino Terminus of Glycoprotein K (gK) Physically Interact with gB
复制标题
DOI：
10.1128/jvi.00298-10
复制
发表时间：
2010-09-01
影响因子：
5.4
通讯作者：
Kousoulas, Konstantin G.
中科院分区：
文献类型：
--
作者：
Chouljenko, Vladimir N.;Iyer, Arun V.;Kousoulas, Konstantin G.
单纯疱疹病毒1型(HSV-1)糖蛋白K(Gk)和UL20蛋白(UL20p)是病毒诱导的细胞融合的严格要求，gk或UL20基因内的突变都会导致广泛的细胞融合(合体形成)。我们已经证明了gk与UL20p形成一个功能性蛋白质复合体，这是HSV-1生命周期中所有gk和UL20p相关功能所必需的。最近，我们发现GK(GKA)的氨基末端82个氨基酸(AA)是表达突变型病毒GB Delta 28合胞体表型所必需的，缺少GB(V.N.Chouljenko，A.V.Iyer，S.Chowdhury，D.V.Chouljenko，K.G.Kousoulas，J.Virol)的28个氨基酸。83：12301-12313,2009年)。提示gk的氨基末端可能直接或间接地与gB和/或其他病毒糖蛋白相互作用。双向免疫共沉淀实验表明，UL20p在感染细胞中与Gb相互作用。此外，GKA多肽与Gb免疫共沉淀，但不与Gd共沉淀。构建了三种重组杆状病毒，分别表达GKA氨基末端82个氨基酸和Gb(30~748 aa)、Gd(1~340 aa)和Gh(1~792 aa)胞外部分。免疫共沉淀实验表明，GKA与Gb和Gh胞外部分发生物理相互作用，但不与Gd相互作用。另外构建了3个表达GKA和截短GBS的重组杆状病毒，分别编码30~154、30~364和30~500个氨基酸。免疫共沉淀实验表明，GKA与所有三种截短的GBS发生物理相互作用。计算机辅助预测GKA在GB上可能的结合位点表明GKA可能主要与GB结构域I相互作用(E.Heldwein，H.Lou，F.C.Bender，G.H.Cohen，R.J.Eisenberg，和S.C.Harrison，Science 313：217-220,2006)。这些结果表明，gk/UL20p蛋白复合体通过直接的物理相互作用调节Gb和Gh的融合特性。
Herpes simplex virus type 1 (HSV-1) glycoprotein K (gK) and the UL20 protein (UL20p) are strictly required for virus-induced cell fusion, and mutations within either the gK or UL20 gene cause extensive cell fusion (syncytium formation). We have shown that gK forms a functional protein complex with UL20p, which is required for all gK and UL20p-associated functions in the HSV-1 life cycle. Recently, we showed that the amino-terminal 82 amino acids (aa) of gK (gKa) were required for the expression of the syncytial phenotype of the mutant virus gB Delta 28 lacking the carboxyl-terminal 28 amino acids of gB (V. N. Chouljenko, A. V. Iyer, S. Chowdhury, D. V. Chouljenko, and K. G. Kousoulas, J. Virol. 83: 12301-12313, 2009). This work suggested that the amino terminus of gK may directly or indirectly interact with gB and/or other viral glycoproteins. Two-way coimmunoprecipitation experiments revealed that UL20p interacted with gB in infected cells. Furthermore, the gKa peptide was coimmunoprecipitated with gB but not gD. Three recombinant baculoviruses were constructed, expressing the amino-terminal 82 aa of gKa together with either the extracellular portion of gB (30 to 748 aa), gD (1 to 340 aa), or gH (1 to 792 aa), respectively. Coimmunoprecipitation experiments revealed that gKa physically interacted with the extracellular portions of gB and gH but not gD. Three additional recombinant baculoviruses expressing gKa and truncated gBs encompassing aa 30 to 154, 30 to 364, and 30 to 500 were constructed. Coimmunoprecipitation experiments showed that gKa physically interacted with all three truncated gBs. Computer-assisted prediction of possible gKa binding sites on gB suggested that gKa may interact predominantly with gB domain I (E. E. Heldwein, H. Lou, F. C. Bender, G. H. Cohen, R. J. Eisenberg, and S. C. Harrison, Science 313: 217-220, 2006). These results imply that the gK/UL20p protein complex modulates the fusogenic properties of gB and gH via direct physical interactions.