Tchénio T, Heidmann T
Unité de Physicochimie et Pharmacologie des Macromolécules Biologigues, Centre National de la Recherche Scientifique URA 147, Institut Gustave Roussy, Villejuif, France.
J Virol. 1995 Feb;69(2):1079-84. doi: 10.1128/JVI.69.2.1079-1084.1995.
Retroviral particles contain a dimer of two genomic RNA molecules, linked by noncovalent intermolecular bonds. Studies by electron microscopy of viral RNA extracted from virions as well as in vitro studies have implicated a sequence, designated the dimer linkage sequence (DLS), in the dimerization process. The DLS has been localized within a short region encompassing the psi packaging sequence, between nucleotides 212 and 563 for the Moloney murine leukemia retrovirus (MoMLV) RNA. In this report, we show that viral RNAs lacking both the DLS and psi packaging sequences--and even an RNA lacking the first 6,537 nucleotides of MoMLV--can assemble within retroviral particles as high-molecular-weight, slow-migrating, heat-sensitive complexes closely related to those observed for wild-type viral RNAs. Furthermore, we show that proviruses of normal structure are generated upon infection of test cells with retroviral particles which contain the DLS/psi-deleted viral RNAs. These observations demonstrate that the DLS and psi packaging sequences are not essential in cis to form a functional RNA complex for reverse transcription and integration.
逆转录病毒颗粒包含由非共价分子间键连接的两个基因组RNA分子的二聚体。对从病毒粒子中提取的病毒RNA进行的电子显微镜研究以及体外研究表明,在二聚化过程中涉及一个被称为二聚体连接序列(DLS)的序列。对于莫洛尼鼠白血病病毒(MoMLV)RNA,DLS已定位在一个短区域内,该区域包含ψ包装序列,位于核苷酸212至563之间。在本报告中,我们表明,既缺乏DLS又缺乏ψ包装序列的病毒RNA,甚至缺乏MoMLV前6537个核苷酸的RNA,都可以在逆转录病毒颗粒内组装成与野生型病毒RNA所观察到的紧密相关的高分子量、慢迁移、热敏感复合物。此外,我们表明,用含有缺失DLS/ψ的病毒RNA的逆转录病毒颗粒感染测试细胞后,会产生正常结构的前病毒。这些观察结果表明,DLS和ψ包装序列对于形成用于逆转录和整合的功能性RNA复合物并非顺式必需的。