Target-cell-derived tRNA-like primers for reverse transcription support retroviral infection at low efficiency.

TY - JOUR

T1 - Target-cell-derived tRNA-like primers for reverse transcription support retroviral infection at low efficiency.

AU - Schmitz, Alexander

AU - Lund, Anders H

AU - Hansen, Anette C

AU - Duch, Mogens

AU - Pedersen, Finn Skou

N1 - Keywords: 3T3 Cells; Animals; Base Sequence; Cell Line; Fibroblasts; Mice; Molecular Sequence Data; Moloney murine leukemia virus; RNA, Transfer; RNA, Transfer, Amino Acyl; RNA, Transfer, Pro; Sequence Alignment; Transcription, Genetic; Virus Replication

PY - 2002

Y1 - 2002

N2 - 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.

AB - 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.

M3 - Journal article

C2 - 12083837

SN - 0042-6822

VL - 297

SP - 68

EP - 77

JO - Virology

JF - Virology

IS - 1

ER -