Old World monkey TRIM5α strongly suppresses human immunodeficiency virus type 1 (HIV-1) replication. A fusion protein comprising cynomolgus macaque (CM) TRIM5 and cyclophilin A (CM TRIMCyp) also potently suppresses HIV-1 replication. However, CM TRIMCyp fails to suppress a mutant HIV-1 that encodes a mutant capsid protein containing a SIVmac239-derived loop between α-helices 4 and 5 (L4/5). There are seven amino acid differences between L4/5 of HIV-1 and SIVmac239. Here, we investigated the minimum numbers of amino acid substitutions that would allow HIV-1 to evade CM TRIMCyp-mediated suppression. We performed random PCR mutagenesis to construct a library of HIV-1 variants containing mutations in L4/5, and then we recovered replication-competent viruses from CD4+ MT4 cells that expressed high levels of CM TRIMCyp. CM TRIMCyp-resistant viruses were obtained after three rounds of selection in MT4 cells expressing CM TRIMCyp and these were found to contain four amino acid substitutions (H87R, A88G, P90D and P93A) in L4/5. We then confirmed that these substitutions were sufficient to confer CM TRIMCyp resistance to HIV-1. In a separate experiment using a similar method, we obtained novel CM TRIM5α-resistant HIV-1 strains after six rounds of selection and rescue. Analysis of these mutants revealed that V86A and G116E mutations in the capsid region conferred partial resistance to CM TRIM5α without substantial fitness cost when propagated in MT4 cells expressing CM TRIM5α. These results confirmed and further extended the previous notion that CM TRIMCyp and CM TRIM5α recognize the HIV-1 capsid in different manners.
AkagiT., OhtsukiY., ShiraishiY., MiyoshiI.1985; Transformation of human fetal thymus and spleen lymphocytes by human T-cell leukemia virus type I. Acta Med Okayama 39:155–159[PubMed]
AndersonJ. L., CampbellE. M., WuX., VandegraaffN., EngelmanA., HopeT. J.2006; Proteasome inhibition reveals that a functional preintegration complex intermediate can be generated during restriction by diverse TRIM5 proteins. J Virol 80:9754–9760 [View Article][PubMed]
BraatenD., FrankeE. K., LubanJ.1996; Cyclophilin A is required for the replication of group M human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus SIV(CPZ)GAB but not group O HIV-1 or other primate immunodeficiency viruses. J Virol 70:4220–4227[PubMed]
ChatterjiU., BobardtM. D., StanfieldR., PtakR. G., PallanschL. A., WardP. A., JonesM. J., StoddartC. A., ScalfaroP., other authors. 2005; Naturally occurring capsid substitutions render HIV-1 cyclophilin A independent in human cells and TRIM-cyclophilin-resistant in owl monkey cells. J Biol Chem 280:40293–40300 [View Article][PubMed]
HowardB. R., VajdosF. F., LiS., SundquistW. I., HillC. P.2003; Structural insights into the catalytic mechanism of cyclophilin A. Nat Struct Biol 10:475–481 [View Article][PubMed]
KirmaierA., WuF., NewmanR. M., HallL. R., MorganJ. S., O'ConnorS., MarxP. A., MeythalerM., GoldsteinS., other authors. 2010; TRIM5 suppresses cross-species transmission of a primate immunodeficiency virus and selects for emergence of resistant variants in the new species. PLoS Biol 8:e1000462 [View Article][PubMed]
KonoK., SongH., ShingaiY., ShiodaT., NakayamaE. E.2008; Comparison of anti-viral activity of rhesus monkey and cynomolgus monkey TRIM5αs against human immunodeficiency virus type 2 infection. Virology 373:447–456 [View Article][PubMed]
KuroishiA., SaitoA., ShingaiY., ShiodaT., NomaguchiM., AdachiA., AkariH., NakayamaE. E.2009; Modification of a loop sequence between α-helices 6 and 7 of virus capsid (CA) protein in a human immunodeficiency virus type 1 (HIV-1) derivative that has simian immunodeficiency virus (SIVmac239) vif and CA α-helices 4 and 5 loop improves replication in cynomolgus monkey cells. Retrovirology 6:70 [View Article][PubMed]
KuroishiA., BozekK., ShiodaT., NakayamaE. E.2010; A single amino acid substitution of the human immunodeficiency virus type 1 capsid protein affects viral sensitivity to TRIM5α. Retrovirology 7:58 [View Article][PubMed]
LiY., KarA. K., SodroskiJ.2009; Target cell type-dependent modulation of human immunodeficiency virus type 1 capsid disassembly by cyclophilin A. J Virol 83:10951–10962 [View Article][PubMed]
MiyamotoT., YokoyamaM., KonoK., ShiodaT., SatoH., NakayamaE. E.2011; A single amino acid of human immunodeficiency virus type 2 capsid protein affects conformation of two external loops and viral sensitivity to TRIM5α. PLoS One 6:e22779 [View Article][PubMed]
NakayamaE. E., MiyoshiH., NagaiY., ShiodaT.2005; A specific region of 37 amino acid residues in the SPRY (B30.2) domain of African green monkey TRIM5α determines species-specific restriction of simian immunodeficiency virus SIVmac infection. J Virol 79:8870–8877 [View Article][PubMed]
NomaguchiM., YokoyamaM., KonoK., NakayamaE. E., ShiodaT., DoiN., FujiwaraS., SaitoA., AkariH., other authors. 2013a; Generation of rhesus macaque-tropic HIV-1 clones that are resistant to major anti-HIV-1 restriction factors. J Virol 87:11447–11461 [View Article][PubMed]
QiM., YangR., AikenC.2008; Cyclophilin A-dependent restriction of human immunodeficiency virus type 1 capsid mutants for infection of nondividing cells. J Virol 82:12001–12008 [View Article][PubMed]
SakuragiJ., UedaS., IwamotoA., ShiodaT.2003; Possible role of dimerization in human immunodeficiency virus type 1 genome RNA packaging. J Virol 77:4060–4069 [View Article][PubMed]
SheehyA. M., GaddisN. C., ChoiJ. D., MalimM. H.2002; Isolation of a human gene that inhibits HIV-1 infection and is suppressed by the viral Vif protein. Nature 418:646–650 [View Article][PubMed]
SongH., NakayamaE. E., YokoyamaM., SatoH., LevyJ. A., ShiodaT.2007; A single amino acid of the human immunodeficiency virus type 2 capsid affects its replication in the presence of cynomolgus monkey and human TRIM5αs. J Virol 81:7280–7285 [View Article][PubMed]
StremlauM., OwensC. M., PerronM. J., KiesslingM., AutissierP., SodroskiJ.2004; The cytoplasmic body component TRIM5α restricts HIV-1 infection in Old World monkeys. Nature 427:848–853 [View Article][PubMed]
Van DammeN., GuatelliJ.2008; HIV-1 Vpu inhibits accumulation of the envelope glycoprotein within clathrin-coated, Gag-containing endosomes. Cell Microbiol 10:1040–1057 [View Article][PubMed]
VeilletteM., BichelK., PawlicaP., FreundS. M., PlourdeM. B., PhamQ. T., Reyes-MorenoC., JamesL. C., BerthouxL.2013; The V86M mutation in HIV-1 capsid confers resistance to TRIM5α by abrogation of cyclophilin A-dependent restriction and enhancement of viral nuclear import. Retrovirology 10:25 [View Article][PubMed]
YlinenL. M., KeckesovaZ., WilsonS. J., RanasingheS., TowersG. J.2005; Differential restriction of human immunodeficiency virus type 2 and simian immunodeficiency virus SIVmac by TRIM5α alleles. J Virol 79:11580–11587 [View Article][PubMed]