%0 Journal Article
%T Assisted Evolution Enables HIV-1 to Overcome a High TRIM5¦Á-Imposed Genetic Barrier to Rhesus Macaque Tropism
%A Steven J. Soll
%A Sam J. Wilson
%A Sebla B. Kutluay
%A Theodora Hatziioannou
%A Paul D. Bieniasz
%J PLOS Pathogens
%D 2013
%I Public Library of Science (PLoS)
%R 10.1371/journal.ppat.1003667
%X Diversification of antiretroviral factors during host evolution has erected formidable barriers to cross-species retrovirus transmission. This phenomenon likely protects humans from infection by many modern retroviruses, but it has also impaired the development of primate models of HIV-1 infection. Indeed, rhesus macaques are resistant to HIV-1, in part due to restriction imposed by the TRIM5¦Á protein (rhTRIM5¦Á). Initially, we attempted to derive rhTRIM5¦Á-resistant HIV-1 strains using two strategies. First, HIV-1 was passaged in engineered human cells expressing rhTRIM5¦Á. Second, a library of randomly mutagenized capsid protein (CA) sequences was screened for mutations that reduced rhTRIM5¦Á sensitivity. Both approaches identified several individual mutations in CA that reduced rhTRIM5¦Á sensitivity. However, neither approach yielded mutants that were fully resistant, perhaps because the locations of the mutations suggested that TRIM5¦Á recognizes multiple determinants on the capsid surface. Moreover, even though additive effects of various CA mutations on HIV-1 resistance to rhTRIM5¦Á were observed, combinations that gave full resistance were highly detrimental to fitness. Therefore, we employed an ¡®assisted evolution¡¯ approach in which individual CA mutations that reduced rhTRIM5¦Á sensitivity without fitness penalties were randomly assorted in a library of viral clones containing synthetic CA sequences. Subsequent passage of the viral library in rhTRIM5¦Á-expressing cells resulted in the selection of individual viral species that were fully fit and resistant to rhTRIM5¦Á. These viruses encoded combinations of five mutations in CA that conferred complete or near complete resistance to the disruptive effects of rhTRIM5¦Á on incoming viral cores, by abolishing recognition of the viral capsid. Importantly, HIV-1 variants encoding these CA substitutions and SIVmac239 Vif replicated efficiently in primary rhesus macaque lymphocytes. These findings demonstrate that rhTRIM5¦Á is difficult to but not impossible to evade, and doing so should facilitate the development of primate models of HIV-1 infection.
%U http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1003667