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TRIM5 alpha drives SIVsmm evolution in rhesus macaques.

ABSTRACT: The antagonistic interaction with host restriction proteins is a major driver of evolutionary change for viruses. We previously reported that polymorphisms of the TRIM5? B30.2/SPRY domain impacted the level of SIVsmm viremia in rhesus macaques. Viremia in macaques homozygous for the non-restrictive TRIM5? allele TRIM5(Q) was significantly higher than in macaques expressing two restrictive TRIM5alpha alleles TRIM5(TFP/TFP) or TRIM5(Cyp/TFP). Using this model, we observed that despite an early impact on viremia, SIVsmm overcame TRIM5? restriction at later stages of infection and that increasing viremia was associated with specific amino acid substitutions in capsid. Two amino acid substitutions (P37S and R98S) in the capsid region were associated with escape from TRIM5(TFP) restriction and substitutions in the CypA binding-loop (GPLPA87-91) in capsid were associated with escape from TRIM5(Cyp). Introduction of these mutations into the original SIVsmE543 clone not only resulted in escape from TRIM5? restriction in vitro but the P37S and R98S substitutions improved virus fitness in macaques with homozygous restrictive TRIM(TFP) alleles in vivo. Similar substitutions were observed in other SIVsmm strains following transmission and passage in macaques, collectively providing direct evidence that TRIM5? exerts selective pressure on the cross-species transmission of SIV in primates.

SUBMITTER: Wu F

PROVIDER: S-EPMC3749954 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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TRIM5 alpha drives SIVsmm evolution in rhesus macaques.

Wu Fan F Kirmaier Andrea A Goeken Robert R Ourmanov Ilnour I Hall Laura L Morgan Jennifer S JS Matsuda Kenta K Buckler-White Alicia A Tomioka Keiko K Plishka Ronald R Whitted Sonya S Johnson Welkin W Hirsch Vanessa M VM

PLoS pathogens 20130822 8

The antagonistic interaction with host restriction proteins is a major driver of evolutionary change for viruses. We previously reported that polymorphisms of the TRIM5α B30.2/SPRY domain impacted the level of SIVsmm viremia in rhesus macaques. Viremia in macaques homozygous for the non-restrictive TRIM5α allele TRIM5(Q) was significantly higher than in macaques expressing two restrictive TRIM5alpha alleles TRIM5(TFP/TFP) or TRIM5(Cyp/TFP). Using this model, we observed that despite an early imp ...[more]

PMID: 23990789

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Project description:UnlabelledTRIM5? proteins are a potent barrier to the cross-species transmission of retroviruses. TRIM5? proteins exhibit an ability to self-associate at many levels, ultimately leading to the formation of protein assemblies with hexagonal symmetry in vitro and cytoplasmic assemblies when expressed in cells. However, the role of these assemblies in restriction, the determinants that mediate their formation, and the organization of TRIM5? molecules within these assemblies have remained unclear. Here we show that ?-helical elements within the Linker2 region of rhesus macaque TRIM5? govern the ability to form cytoplasmic assemblies in cells and restrict HIV-1 infection. Mutations that reduce ?-helix formation by the Linker2 region disrupt assembly and restriction. More importantly, mutations that enhance the ?-helical content of the Linker2 region, relative to the wild-type protein, also exhibit an increased ability to form cytoplasmic assemblies and restrict HIV-1 infection. Molecular modeling of the TRIM5? dimer suggests a model in which ?-helical elements within the Linker2 region dock to ?-helices of the coiled-coil domain, likely establishing proper orientation and spacing of protein domains necessary for assembly and restriction. Collectively, these studies provide critical insight into the determinants governing TRIM5? assembly and restriction and demonstrate that the antiviral potency of TRIM5? proteins can be significantly increased without altering the affinity of SPRY/capsid binding.ImportanceMany members of the tripartite motif (TRIM) family of proteins act as restriction factors that directly inhibit viral infection and activate innate immune signaling pathways. Another common feature of TRIM proteins is the ability to form protein assemblies in the nucleus or the cytoplasm. However, the determinants in TRIM proteins required for assembly and the degree to which assembly affects TRIM protein function have been poorly understood. Here we show that alpha helices in the Linker2 (L2) region of rhesus TRIM5? govern assembly and restriction of HIV-1 infection. Helix-disrupting mutations disrupt the assembly and restriction of HIV-1, while helix-stabilizing mutations enhance assembly and restriction relative to the wild-type protein. Circular dichroism analysis suggests that that the formation of this helical structure is supported by intermolecular interactions with the coiled-coil (CC) domain in the CCL2 dimer. These studies reveal a novel mechanism by which the antiviral activity of TRIM5? proteins can be regulated and provide detailed insight into the assembly determinants of TRIM family proteins.

Project description: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? protein (rhTRIM5?). Initially, we attempted to derive rhTRIM5?-resistant HIV-1 strains using two strategies. First, HIV-1 was passaged in engineered human cells expressing rhTRIM5?. Second, a library of randomly mutagenized capsid protein (CA) sequences was screened for mutations that reduced rhTRIM5? sensitivity. Both approaches identified several individual mutations in CA that reduced rhTRIM5? sensitivity. However, neither approach yielded mutants that were fully resistant, perhaps because the locations of the mutations suggested that TRIM5? recognizes multiple determinants on the capsid surface. Moreover, even though additive effects of various CA mutations on HIV-1 resistance to rhTRIM5? 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? 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?-expressing cells resulted in the selection of individual viral species that were fully fit and resistant to rhTRIM5?. These viruses encoded combinations of five mutations in CA that conferred complete or near complete resistance to the disruptive effects of rhTRIM5? on incoming viral cores, by abolishing recognition of the viral capsid. Importantly, HIV-1 variants encoding these CA substitutions and SIV(mac239) Vif replicated efficiently in primary rhesus macaque lymphocytes. These findings demonstrate that rhTRIM5? is difficult to but not impossible to evade, and doing so should facilitate the development of primate models of HIV-1 infection.

Dataset Information

TRIM5 alpha drives SIVsmm evolution in rhesus macaques.

Publications

TRIM5 alpha drives SIVsmm evolution in rhesus macaques.

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