Target-cell-derived tRNA-like primers for reverse transcription support retroviral infection at low efficiency.
Reverse transcription of a retroviral genome takes place in the cytoplasm of an infected cell by a process primed by a producer-cell-derived tRNA annealed to an 18-nucleotide primer-binding site (PBS). By an assay involving primer complementation of PBS-mutated vectors we analyzed whether tRNA primers derived from the target cell can sustain reverse transcription during murine leukemia virus (MLV) infection. Transduction efficiencies were 4-5 orders of magnitude below those of comparable producer-cell complementations. However, successful usage of a target-cell-derived tRNA primer was proven by cases of correction of single mismatches between Akv-MLV vectors and complementary tRNA primers toward the primer sequence in the integrated vector. Thus, target-cell-derived tRNA-like primers are able to initiate first-strand cDNA synthesis and plus-strand transfer leading to a complete provirus, suggesting that endogenous tRNAs from the infected cell may also have access to the intracellular viral complex at that step of the replication cycle.
Schmitz
Alexander
A
Department of Molecular and Structural Biology, Aarhus University, C. F. Moellers Allé, Building 130, Denmark.
Lund
Anders H
AH
Hansen
Anette C
AC
Duch
Mogens
M
Pedersen
Finn Skou
FS
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005347:Fibroblasts / Q000737:chemistry / Q000821:virology; D051379:Mice; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics / Q000502:physiology*; D012343:RNA, Transfer / Q000502:physiology*; D012346:RNA, Transfer, Amino Acyl / Q000502:physiology; D012361:RNA, Transfer, Pro / Q000502:physiology; D016415:Sequence Alignment; D014158:Transcription, Genetic / Q000502:physiology; D014779:Virus Replication
D012346:RNA, Transfer, Amino Acyl; D012361:RNA, Transfer, Pro; D012343:RNA, Transfer