Fusion of cyclophilin A to Fv1 enables cyclosporine-sensitive restriction of human and feline immunodeficiency viruses.
TRIM5alpha is a potent intracellular antiviral restriction factor governing species-specific retroviral replication. In the New World species owl monkey the coding region for the viral binding B30.2 domain of TRIM5alpha has been replaced by a cyclophilin A (CypA) pseudogene by retrotransposition. The resultant TRIM5-CypA fusion protein restricts human immunodeficiency virus type 1 (HIV-1), as well as feline immunodeficiency virus (FIV), by recruitment of the CypA domain to the incoming viral capsids. Infectivity is rescued by agents such as cyclosporine that disrupt CypA binding to its substrates. Mice encode an antiviral restriction factor called Fv1 (for Friend virus susceptibility gene 1), which is active against murine leukemia virus and related to endogenous gag sequences. Here we show that fusing CypA to Fv1 generates a restriction factor with the antiviral specificity of TRIMCyp but the antiviral properties of Fv1. Like TRIMCyp, Fv1-Cyp restricts HIV-1 and FIV and is sensitive to inhibition by cyclosporine. TRIM5alpha is known to have a short half-life and block infectivity before viral reverse transcription. We show that Fv1-Cyp has a long half-life and blocks after reverse transcription, suggesting that its longer half-life gives the restricted virus the opportunity to synthesize DNA, leading to a later block to infection. This notion is supported by the observation that infectivity of Fv1-Cyp restricted virus can be rescued by cyclosporine for several hours after infection, whereas virus restricted by TRIMCyp is terminally restricted after around 40 min. Intriguingly, the Fv1-Cyp-restricted HIV-1 generates closed circular viral DNA, suggesting that the restricted virus complex enters the nucleus.
Schaller
Torsten
T
MRC Centre for Medical Molecular Virology, Department of Infection, Royal Free and University College London Medical School, 46 Cleveland Street, London, United Kingdom.
Ylinen
Laura M J
LM
Webb
Benjamin L J
BL
Singh
Shalene
S
Towers
Greg J
GJ
D000818:Animals; D016645:Aotidae; D002213:Capsid / Q000276:immunology / Q000378:metabolism; D002415:Cats; D002460:Cell Line; D021984:Cyclophilin A / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics; D016089:Immunodeficiency Virus, Feline / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D011506:Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D048348:Reverse Transcription / Q000235:genetics / Q000276:immunology; D053586:Virus Internalization
D004279:DNA, Viral; C103189:Fv1 protein, mouse; D011506:Proteins; D011993:Recombinant Fusion Proteins; D021984:Cyclophilin A