The S40 residue in HIV-1 Gag p6 impacts local and distal budding determinants, revealing additional late domain activities

Retrovirology

Carol A Carter²  Harold Burger⁶  Kathryn Anastos⁴  Michael Powell⁷  Chaoping Chen¹  Binshan Shi⁵  Barbara Weiser⁶  Kimdar Sherefa Kemal³  Lorna Ehrlich²  Mahfuz Khan⁷  Min-Huei Chen²  Susan M Watanabe² 

[1] Colorado State University, Fort Collins, CO 80523, USA;Department of Molecular Genetics & Microbiology, Stony Brook University, Life Sciences Bldg. Rm 248, Stony Brook, NY 11794-5222, USA;Current address: Albert Einstein College of Medicine, Bronx, NY 10461, USA;Albert Einstein College of Medicine, Bronx, NY 10461, USA;Current address: Albany College of Pharmacy and Health Sciences, Albany, NY 12208, USA;Current address: University of California Davis School of Medicine and Sacramento VA Medical Center, Sacramento, CA 95817, USA;Department of Microbiology & Immunology, Morehouse School of Medicine, Atlanta, GA 30310, USA

关键词: Viral budding;    Protease;    Viral particle maturation;    CA NTD;    Nedd4;    Tsg101;    Alix;    HIV-1;   

Others  :  806085
DOI  :  10.1186/1742-4690-10-143

 received in 2013-05-10, accepted in 2013-11-11,  发布年份 2013

【 摘 要 】

Background

HIV-1 budding is directed primarily by two motifs in Gag p6 designated as late domain-1 and −2 that recruit ESCRT machinery by binding Tsg101 and Alix, respectively, and by poorly characterized determinants in the capsid (CA) domain. Here, we report that a conserved Gag p6 residue, S40, impacts budding mediated by all of these determinants.

Results

Whereas budding normally results in formation of single spherical particles ~100 nm in diameter and containing a characteristic electron-dense conical core, the substitution of Phe for S40, a change that does not alter the amino acids encoded in the overlapping pol reading frame, resulted in defective CA-SP1 cleavage, formation of strings of tethered particles or filopodia-like membrane protrusions containing Gag, and diminished infectious particle formation. The S40F-mediated release defects were exacerbated when the viral-encoded protease (PR) was inactivated or when L domain-1 function was disrupted or when budding was almost completely obliterated by the disruption of both L domain-1 and −2. S40F mutation also resulted in stronger Gag-Alix interaction, as detected by yeast 2-hybrid assay. Reducing Alix binding by mutational disruption of contact residues restored single particle release, implicating the perturbed Gag-Alix interaction in the aberrant budding events. Interestingly, introduction of S40F partially rescued the negative effects on budding of CA NTD mutations EE75,76AA and P99A, which both prevent membrane curvature and therefore block budding at an early stage.

Conclusions

The results indicate that the S40 residue is a novel determinant of HIV-1 egress that is most likely involved in regulation of a critical assembly event required for budding in the Tsg101-, Alix-, Nedd4- and CA N-terminal domain affected pathways.

【 授权许可】

   
2013 Watanabe et al.; licensee BioMed Central Ltd.

【 图 表 】

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