Bogerd H, Greene W C
Department of Medicine, Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710.
J Virol. 1993 May;67(5):2496-502. doi: 10.1128/JVI.67.5.2496-2502.1993.
Human T-cell leukemia virus type I (HTLV-I) Rex and human immunodeficiency virus type 1 (HIV-1) Rev are essential gene products required for the replication of these two pathogenic human retroviruses. Both Rex and Rev act at a posttranscriptional level by binding to highly structured RNA-response elements, the Rex-response element in HTLV-I and the Rev-response element in HIV-1. Using a sensitive in vivo assay of protein-protein interaction, we now demonstrate that the HTLV-I Rex and HIV-1 Rev proteins readily form homomultimeric complexes in the absence of their cognate RNA-response elements yet fail to form heteromultimeric complexes with each other. Dominant negative mutations have been identified in both the rex and rev genes which presumably specify a critical activation or effector domain in each of these viral transactivators. Surprisingly, these dominant negative mutants of Rex and Rev fail to interact in vivo. These findings raise the possibility that the binding of nonfunctional monomers rather than functional multimers underlies the transdominant phenotype of these Rex and Rev mutants. Further, it seems likely that the assembly of functional and stable multimers of Rex and Rev in vivo may depend not only on the intrinsic multimerization domains of these proteins but also on the binding of a bridging cellular cofactor to the related activation domains present in each viral transactivator.
人类嗜T淋巴细胞病毒I型(HTLV-I)的Rex蛋白和人类免疫缺陷病毒1型(HIV-1)的Rev蛋白是这两种致病性人类逆转录病毒复制所必需的基因产物。Rex和Rev都通过与高度结构化的RNA反应元件结合而在转录后水平发挥作用,HTLV-I中的Rex反应元件和HIV-1中的Rev反应元件。利用一种灵敏的体内蛋白质-蛋白质相互作用检测方法,我们现在证明,在没有其同源RNA反应元件的情况下,HTLV-I的Rex蛋白和HIV-1的Rev蛋白很容易形成同多聚体复合物,但彼此之间不能形成异多聚体复合物。在rex和rev基因中都已鉴定出显性负性突变,这些突变大概在每种病毒反式激活因子中指定了一个关键的激活或效应结构域。令人惊讶的是,Rex和Rev的这些显性负性突变体在体内无法相互作用。这些发现增加了一种可能性,即这些Rex和Rev突变体的反式显性表型的基础是无功能单体的结合而非功能多聚体的结合。此外,Rex和Rev在体内形成功能稳定多聚体的组装似乎不仅取决于这些蛋白质的内在多聚化结构域,还取决于一种桥接细胞辅因子与每种病毒反式激活因子中存在的相关激活结构域的结合。