Schwartzberg P, Colicelli J, Gordon M L, Goff S P
J Virol. 1984 Mar;49(3):918-24. doi: 10.1128/JVI.49.3.918-924.1984.
We have constructed a series of deletion mutations in the p30 and p10 domains of the gag gene of Moloney murine leukemia virus. Mutants with deletions in P30 were completely defective in virion particle production even though an altered gag precursor protein is synthesized. This domain is apparently critical for particle formation. A mutant in P10 was able to release virion particles into the medium, and low levels of reverse transcriptase activity could be detected in these virions. To explore the effects of these mutations on the utilization of the gag-pol precursor, we have introduced these mutants into cells already releasing defective particles from an endogenous provirus which directs the synthesis of gag gene products and not pol gene products. The P10 mutant was capable of providing pol function as judged by the incorporation of high levels of reverse transcriptase into the particles and complete complementation for XC plaque formation. In contrast, the mutants in P30 were negative in this complementation test. Thus, those gag mutants which were unable on their own to assemble virion particles were also unable to contribute the gag-pol precursor to these particles. These mutations are the first to be mapped to the gag region which affect pol function, suggesting that the gag-pol precursor must be assembled before pol is functionally separated from the gag domain. The concordance of the effects of different mutations on both particle formation and gag-pol utilization suggests that similar domains of gag (namely, domains in the P30 region) are needed for these two processes.
我们构建了莫洛尼鼠白血病病毒gag基因p30和p10结构域的一系列缺失突变体。p30缺失的突变体在病毒粒子产生方面完全缺陷,尽管合成了一种改变的gag前体蛋白。该结构域显然对粒子形成至关重要。p10的一个突变体能够将病毒粒子释放到培养基中,并且在这些病毒粒子中可以检测到低水平的逆转录酶活性。为了探究这些突变对gag-pol前体利用的影响,我们将这些突变体引入已经从内源性前病毒释放缺陷粒子的细胞中,该内源性前病毒指导gag基因产物而非pol基因产物的合成。根据高水平逆转录酶掺入粒子以及对XC噬斑形成的完全互补作用判断,p10突变体能够提供pol功能。相比之下,p30的突变体在这种互补试验中呈阴性。因此,那些自身无法组装病毒粒子的gag突变体也无法为这些粒子提供gag-pol前体。这些突变是首次定位到影响pol功能的gag区域,表明gag-pol前体必须在pol从gag结构域功能分离之前组装。不同突变对粒子形成和gag-pol利用的影响的一致性表明,这两个过程需要gag的相似结构域(即p30区域的结构域)。