title: The contribution of RING and B-box 2 domains to retroviral restriction mediated by monkey TRIM5alpha.
abstract: TRIM5alpha is a cytoplasmic protein that mediates a post-entry block to infection by some retroviruses. TRIM5alpha contains a tripartite motif (TRIM), which includes RING, B-box 2, and coiled-coil domains, and a C-terminal B30.2 (SPRY) domain. We investigated the contribution of the RING and B-box 2 domains to the antiretroviral activity of rhesus monkey TRIM5alpha (TRIM5alpharh), which potently restricts infection by human immunodeficiency virus, type 1 (HIV-1) and simian immunodeficiency virus of African green monkeys (SIVagm). Disruption of the RING domain caused mislocalization of TRIM5alpharh so that the cytoplasmic level of the protein was decreased compared with that of the wild-type protein. Nonetheless, partial ability to restrict HIV-1 and SIVagm was retained by the RING domain mutants. By contrast, although TRIM5alpharh mutants with disrupted B-box 2 domains were efficiently expressed and correctly localized to the cytoplasm, antiretroviral activity was absent. The B-box 2 mutants colocalized and associated with wild-type TRIM5alpharh and exerted dominant-negative effects on the antiretroviral activity of the wild-type protein. Taken together with other data, these results indicate that functionally defective TRIM5alpharh molecules that retain a coiled coil can act as dominant-negative inhibitors of wild-type TRIM5alpharh function. The RING domain of TRIM5alpharh is not absolutely required for retrovirus restriction but can influence cytoplasmic levels of the protein and thus indirectly alter function. The B-box 2 domain, by contrast, appears to be essential for efficient retrovirus restriction.
authors.affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
authors.forename: Hassan
authors.initials: H
authors.lastname: Javanbakht
authors.forename: Felipe
authors.initials: F
authors.lastname: Diaz-Griffero
authors.forename: Matthew
authors.initials: M
authors.lastname: Stremlau
authors.forename: Zhihai
authors.initials: Z
authors.lastname: Si
authors.forename: Joseph
authors.initials: J
authors.lastname: Sodroski
mesh_terms: D000818:Animals; D015497:HIV-1 / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D007113:Immunity, Innate; D008253:Macaca mulatta; D009154:Mutation; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D012190:Retroviridae / Q000276:immunology*; D015302:Simian Immunodeficiency Virus / Q000276:immunology; D014162:Transfection; D044767:Ubiquitin-Protein Ligases; D016335:Zinc Fingers
chemical_list: D011506:Proteins; C482587:TRIM5(alpha) protein, rhesus monkey; D044767:Ubiquitin-Protein Ligases