期刊
VIRUSES-BASEL
卷 13, 期 5, 页码 -出版社
MDPI
DOI: 10.3390/v13050903
关键词
retrovirus; RSV; RNA; dimerization; microscopy; FISH
类别
资金
- NIH [R01 CA76534, R01 GM139392, F30 CA214010, 1S10OD010756-01A1]
- Penn State College of Medicine
- Summer Bridge Award
The mechanism of genomic RNA dimer formation in Rous sarcoma virus is more similar to murine leukemia virus (MLV) than human immunodeficiency virus (HIV-1). This study revealed that genomic RNA heterodimers can form in the nucleus, cytoplasm, and at the plasma membrane, but RSV preferentially packages genome homodimers into virus particles.
Retroviruses are unique in that they package their RNA genomes as non-covalently linked dimers. Failure to dimerize their genomes results in decreased infectivity and reduced packaging of genomic RNA into virus particles. Two models of retrovirus genome dimerization have been characterized: in murine leukemia virus (MLV), genomic RNA dimerization occurs co-transcriptionally in the nucleus, resulting in the preferential formation of genome homodimers; whereas in human immunodeficiency virus (HIV-1), genomic RNA dimerization occurs in the cytoplasm and at the plasma membrane, with a random distribution of heterodimers and homodimers. Although in vitro studies have identified the genomic RNA sequences that facilitate dimerization in Rous sarcoma virus (RSV), in vivo characterization of the location and preferences of genome dimerization has not been performed. In this study, we utilized three single molecule RNA imaging approaches to visualize genome dimers of RSV in cultured quail fibroblasts. The formation of genomic RNA heterodimers within cells was dependent on the presence of the dimerization initiation site (DIS) sequence in the L3 stem. Subcellular localization analysis revealed that heterodimers were present the nucleus, cytoplasm, and at the plasma membrane, indicating that genome dimers can form in the nucleus. Furthermore, single virion analysis revealed that RSV preferentially packages genome homodimers into virus particles. Therefore, the mechanism of RSV genomic RNA dimer formation appears more similar to MLV than HIV-1.
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