SEQUENCE REQUIREMENTS FOR REV MULTIMERIZATION IN-VIVO
SEQUENCE REQUIREMENTS FOR REV MULTIMERIZATION IN-VIVO
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DOI：
10.1006/viro.1994.1334
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发表时间：
1994-07-01
期刊：
影响因子：
3.7
通讯作者：
CULLEN, BR
中科院分区：
文献类型：
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作者：
MADORE, SJ;TILEY, LS;CULLEN, BR
人类免疫缺陷病毒1型（HIV-1）Rev蛋白的多聚化被认为是其生物活性的关键。然而，Rev体内多聚化的精确蛋白质序列要求，以及多聚化是否由特异性RNA结合促进或反之亦然，仍然存在争议。在这份报告中，我们描述了一种灵敏的体内检测HIV-1 Rev在其同源RRE主要RNA结合位点上的多聚化。使用这种方法，我们证明了一个完整的Rev精氨酸丰富的结构域，而特异性RNA结合的关键，是对RRE的多聚化。引入到Rev序列中的突变在该基本结构域的侧翼在体内产生部分多聚化表型，即使已知这些突变在体外阻断Rev多聚化。类似地，突变引入到富含亮氨酸的激活结构域的Rev，这似乎没有影响在体外多聚化，也显着抑制多聚化的Rev的RRE在体内。总之，这些数据似乎与多聚Rev：RRE核糖核蛋白复合物的体内形成由RRE RNA底物促进的假设一致，并且如Bogerd和格林（J.Virol. 67，2496-2502，1993），通过可能与多个Rev活化结构域相互作用的Rev的细胞辅因子桥接。(C)1994年出版社出版。
Multimerization of the human immunodeficiency virus type 1 (HIV-1) Rev protein is believed to be critical to its biological activity. However, the precise protein sequence requirements for Rev multimerization in vivo, and whether multimerization is facilitated by specific RNA binding or vice versa, has remained controversial. In this report, we describe a sensitive in vivo assay for the multimerization of HIV-1 Rev on its cognate RRE primary RNA binding site. Using this assay, we demonstrate that an intact Rev arginine-rich domain, while critical to specific RNA binding, is dispensable for multimerization on the RRE. Mutations introduced into Rev sequences that flank this basic domain produce a partial multimerization phenotype in vivo even though these mutations are known to block Rev multimerization in vitro. Similarly, mutations introduced into the leucine-rich activation domain of Rev, which appear to have no effect on in vitro multimerization, also markedly inhibit multimerization of Rev on the RRE in vivo. Overall, these data appear consistent with the hypothesis that in vivo formation of the multimeric Rev:RRE ribonucleoprotein complex is facilitated by both the RRE RNA substrate and, as first proposed by Bogerd and Greene (J. Virol. 67, 2496-2502, 1993), by bridging by a cellular cofactor for Rev that likely interacts with multiple Rev activation domains. (C) 1994 Academic Press, Inc.