期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 295, 期 42, 页码 14391-14401出版社
ELSEVIER
DOI: 10.1074/jbc.RA120.014835
关键词
human immunodeficiency virus; HIV; virus assembly; mass spectrometry; MS; RNA binding protein; RNA; RNA-protein interaction; native mass spectrometry; genomic RNA packaging; Gag; Gag p6 domain
资金
- National Institutes of Health [P41 GM128577, RO1 AI153216, U54 AI150472, T32 GM118291]
- Intramural Research Program of the Center for Cancer Research, NCI, National Institutes of Health
- Intramural AIDS Targeted Antiviral Therapy Program
The HIV-1 Gag protein is responsible for genomic RNA (gRNA) packaging and immature viral particle assembly. Although the presence of gRNA in virions is required for viral infectivity, in its absence, Gag can assemble around cellular RNAs and form particles resembling gRNA-containing particles. When gRNA is expressed, it is selectively packaged despite the presence of excess host RNA, but how it is selectively packaged is not understood. Specific recognition of a gRNA packaging signal (Psi) has been proposed to stimulate the efficient nucleation of viral assembly. However, the heterogeneity of Gag-RNA interactions renders capturing this transient nucleation complex using traditional structural biology approaches challenging. Here, we used native MS to investigate RNA binding of wild-type (WT) Gag and Gag lacking the p6 domain (Gag Delta p6). Both proteins bind to Psi RNA primarily as dimers, but to a control RNA primarily as monomers. The dimeric complexes on Psi RNA require an intact dimer interface within Gag. Gag Delta p6 binds to Psi RNA with high specificity in vitro and also selectively packages gRNA in particles produced in mammalian cells. These studies provide direct support for the idea that Gag binding to Psi specifically promotes nucleation of Gag-Gag interactions at the early stages of immature viral particle assembly in a p6-independent manner.
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