Lama J, Trono D
Infectious Disease Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037, USA.
J Virol. 1998 Feb;72(2):1671-6. doi: 10.1128/JVI.72.2.1671-1676.1998.
The matrix (MA) protein of human immunodeficiency virus type 1 (HIV-1) plays a critical role in virion morphogenesis and fulfills important functions during the early steps of infection. In an effort to identify cellular partners of MA, a Saccharomyces cerevisiae two-hybrid screen was utilized. A specific interaction between MA and HO3, a putative histidyl-tRNA synthetase, was demonstrated in this system. HO3-specific mRNA was detected in several tissues relevant for HIV infection, such as spleen, thymus, and peripheral blood lymphocytes, as well as in a number of T-lymphoid-cell lines. The binding of MA to HO3 was confirmed in transfected cells by coimmunoprecipitation. This interaction was abrogated by replacing two lysine residues at positions 26 and 27 of MA by threonine (MA(KK27TT)). HO3 localized both to the cytoplasm and to the nucleus of acutely transfected 293T cells. When overexpressed in HIV-1-producing cells, HO3 was incorporated into wild-type virions but not in ones containing the dilysine-mutated variant of MA. Correspondingly, overexpression of HO3 in virus producer cells enhanced the infectivity of wild-type but not MA(KK27AA) HIV-1 particles. The stimulating effect of HO3 was independent from the presence of Envelope, Vpr, or Vpu. Taken together, these results suggest that HO3, through its recognition of MA, plays a role in the life cycle of HIV-1.
人类免疫缺陷病毒1型(HIV-1)的基质(MA)蛋白在病毒体形态发生中起关键作用,并在感染早期阶段履行重要功能。为了鉴定MA的细胞伴侣,利用了酿酒酵母双杂交筛选。在该系统中证明了MA与假定的组氨酰-tRNA合成酶HO3之间存在特异性相互作用。在与HIV感染相关的几种组织中,如脾脏、胸腺和外周血淋巴细胞,以及一些T淋巴细胞系中检测到HO3特异性mRNA。通过共免疫沉淀在转染细胞中证实了MA与HO3的结合。将MA第26和27位的两个赖氨酸残基替换为苏氨酸(MA(KK27TT))可消除这种相互作用。HO3定位于急性转染的293T细胞的细胞质和细胞核。当在产生HIV-1的细胞中过表达时,HO3被整合到野生型病毒体中,但不整合到含有MA双赖氨酸突变变体的病毒体中。相应地,在病毒产生细胞中过表达HO3可增强野生型HIV-1颗粒的感染性,但不能增强MA(KK27AA) HIV-1颗粒的感染性。HO3的刺激作用与包膜、Vpr或Vpu的存在无关。综上所述,这些结果表明HO3通过识别MA在HIV-1的生命周期中发挥作用。