Houzet Laurent, Battini Jean Luc, Bernard Eric, Thibert Valerie, Mougel Marylène
Institut de Genetique Moleculaire, UMR5555 CNRS, Montpellier, 4 Boulevard Henri IV, CS89508, 34960 Montpellier, France.
EMBO J. 2003 Sep 15;22(18):4866-75. doi: 10.1093/emboj/cdg450.
Replication of simple retroviruses depends on the recruitment of a single large primary transcript toward splicing, transport/packaging and translation regulations. In this respect, we studied the novel SD' 4.4 kb RNA of murine leukemia retroviruses (MLV) which results from alternative splicing of the primary transcript. We showed that SD' RNA was required for optimal replication since expression of a pre-spliced SD' RNA trans-complemented the impaired infectivity of a SD'-defective mutant. We monitored the fate of this novel transcript throughout early and late events of the viral life cycle. SD' RNA was specifically incorporated into virions demonstrating that the unspliced RNA was not the unique viral RNA present in virions. Furthermore, SD' RNA was reverse transcribed and its DNA copy integrated into the host genome, thus constituting a new splice donor-associated retroelement (SDARE) in infected cells. Finally, we showed that SD' mRNA encoded a 50 kDa polyprotein, and to a lower extent an additional 60 kDa polyprotein, which harbored Gag and integrase domains.
简单逆转录病毒的复制依赖于将单一的大型初级转录本招募至剪接、运输/包装及翻译调控过程。在这方面,我们研究了鼠白血病逆转录病毒(MLV)的新型4.4 kb SD' RNA,它是初级转录本选择性剪接的产物。我们发现SD' RNA是最佳复制所必需的,因为预剪接的SD' RNA的表达可反式互补SD'缺陷型突变体受损的感染性。我们在病毒生命周期的早期和晚期事件中监测了这种新型转录本的命运。SD' RNA被特异性地包装进病毒粒子,这表明未剪接的RNA并非病毒粒子中唯一存在的病毒RNA。此外,SD' RNA被逆转录,其DNA拷贝整合到宿主基因组中,从而在感染细胞中构成一种新的与剪接供体相关的逆转录元件(SDARE)。最后,我们表明SD' mRNA编码一种50 kDa的多蛋白,在较低程度上还编码另一种60 kDa的多蛋白,后者含有Gag和整合酶结构域。