Selection of retroviral reverse transcription primer is coordinated with tRNA biogenesis.
Initiation of retrovirus reverse transcription requires the selection of a tRNA primer from the intracellular milieu. To investigate the features of primer selection, a human immunodeficiency virus type 1 (HIV-1) and a murine leukemia virus (MuLV) were created that require yeast tRNA(Phe) to be supplied in trans for infectivity. Wild-type yeast tRNA(Phe) expressed in mammalian cells was transported to the cytoplasm and aminoacylated. In contrast, tRNA(Phe) without the D loop (tRNA(Phe)D(-)) was retained within the nucleus and did not complement infectivity of either HIV-1 or MuLV; however, infectivity was restored when tRNA(Phe)D(-) was directly transfected into the cytoplasm of cells. A tRNA(Phe) mutant (tRNA(Phe)UUA) that did not have the capacity to be aminoacylated was transported to the cytoplasm and did complement infectivity of both HIV-1 and MuLV, albeit at a level less than that with wild-type tRNA(Phe). Collectively, our results demonstrate that the tRNA primer captured by HIV-1 and MuLV occurs after nuclear export of tRNA and supports a model in which primer selection for retroviruses is coordinated with tRNA biogenesis at the intracellular site of protein synthesis.
Kelly
Nathan J
NJ
Department of Microbiology, University of Alabama at Birmingham, 35294, USA.
Palmer
Matthew T
MT
Morrow
Casey D
CD
D001483:Base Sequence; D002460:Cell Line; D005816:Genetic Complementation Test; D006678:HIV / Q000235:genetics* / Q000502:physiology; D006801:Humans; D009154:Mutation; D009690:Nucleic Acid Conformation; D012313:RNA; D012360:RNA, Transfer, Phe / Q000096:biosynthesis* / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014158:Transcription, Genetic; D014779:Virus Replication
C034477:RNA primers; D012360:RNA, Transfer, Phe; D012313:RNA