Identification of residues within the L2 region of rhesus TRIM5alpha that are required for retroviral restriction and cytoplasmic body localization.
The intracellular restriction factor TRIM5alpha, inhibits infection by numerous retroviruses in a species-specific manner. The best characterized example of this restriction is the TRIM5alpha protein from rhesus macaques (rhTRIM5alpha), which potently inhibits HIV-1 infection. TRIM5alpha localizes to cytoplasmic assemblies of protein referred to as cytoplasmic bodies, though the role that these bodies play in retroviral restriction is unclear. We employed a series of truncation mutants to identify a discrete region, located within the Linker2 region connecting the coiled-coil and B30.2/PRYSPRY domains of TRIM5alpha, which is required for cytoplasmic body localization. Deletion of this region in the context of full-length rhTRIM5alpha abrogates cytoplasmic body localization. Alanine mutagenesis of the residues in this region identifies two stretches of amino acids that are required for both cytoplasmic body localization and retroviral restriction. This work suggests that the determinants that mediate TRIM5alpha localization to cytoplasmic bodies play a requisite role in retroviral restriction.
Sastri
Jaya
J
Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University Chicago, Maywood IL, USA.
O'Connor
Christopher
C
Danielson
Cindy M
CM
McRaven
Michael
M
Perez
Patricio
P
Diaz-Griffero
Felipe
F
Campbell
Edward M
EM
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D003593:Cytoplasm; D004274:DNA, Recombinant; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D008253:Macaca mulatta; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D017434:Protein Structure, Tertiary; D021381:Protein Transport
D002352:Carrier Proteins; D004274:DNA, Recombinant