Human T-cell leukemia virus type 1 uses a specific tRNAProisodecoder to prime reverse transcription

Abstract

Human T-cell leukemia virus type 1 (HTLV-1) is the only oncogenic human retrovirus discovered to date. All retroviruses are believed to use a host cell tRNA to prime reverse transcription (RT). In HTLV-1, the primer-binding site (PBS) in the genomic RNA is complementary to the 3′ 18 nucleotides (nt) of human tRNAPro. The human genome encodes 20 cytoplasmic tRNAProgenes representing seven isodecoders, all of which share the same 3′ 18 nt sequence but vary elsewhere. Whether all tRNAProisodecoders are used to prime RT in cells is unknown. A previous study showed that a 3′ 18 nt tRNAPro-derived fragment (tRFPro) is packaged into HTLV-1 particles and can serve as an RT primer in vitro. The role of this tRNA fragment in the viral life cycle is unclear. In retroviruses, N1-methylation of the tRNA primer at position A58 (m1A) is essential for successful plus-strand transfer. Using primer-extension assays performed in chronically HTLV-1-infected cells, we found that A58 of tRNAProis m1A-modified, implying that full-length tRNAProis capable of facilitating successful plus-strand transfer. Analysis of HTLV-1 RT primer extension products indicated that full-length tRNAProis likely to be the primer. To determine which tRNAProisodecoder is used as the RT primer, we sequenced the minus-strand strong-stop RT product containing the intact tRNA primer and established that HTLV-1 primes RT using a specific tRNAProUGG isodecoder. Further studies are required to understand how this primer is annealed to the highly structured HTLV-1 PBS and to investigate the role of tRFProin the viral life cycle.