Extended minus-strand DNA as template for R-U5-mediated second-strand transfer in recombinational rescue of primer binding site-modified retroviral vectors.

TY - JOUR

T1 - Extended minus-strand DNA as template for R-U5-mediated second-strand transfer in recombinational rescue of primer binding site-modified retroviral vectors.

AU - Mikkelsen, J G

AU - Lund, Anders Henrik

AU - Dybkaer, K

AU - Duch, M

AU - Pedersen, F S

N1 - Keywords: 3T3 Cells; Animals; Base Sequence; Binding Sites; Cell Transformation, Viral; DNA Primers; DNA, Single-Stranded; DNA, Viral; Genetic Vectors; Leukemia Virus, Murine; Mice; Molecular Sequence Data; Recombination, Genetic; Repetitive Sequences, Nucleic Acid; Ribonucleoprotein, U5 Small Nuclear; Templates, Genetic; Virus Replication

PY - 1998

Y1 - 1998

N2 - We have previously demonstrated recombinational rescue of primer binding site (PBS)-impaired Akv murine leukemia virus-based vectors involving initial priming on endogenous viral sequences and template switching during cDNA synthesis to obtain PBS complementarity in second-strand transfer of reverse transcription (Mikkelsen et al., J. Virol. 70:1439-1447, 1996). By use of the same forced recombination system, we have now found recombinant proviruses of different structures, suggesting that PBS knockout vectors may be rescued through initial priming on endogenous virus RNA, read-through of the mutated PBS during minus-strand synthesis, and subsequent second-strand transfer mediated by the R-U5 complementarity of the plus strand and the extended minus-strand DNA acceptor template. Mechanisms for R-U5-mediated second-strand transfer and its possible role in retrovirus replication and evolution are discussed.

AB - We have previously demonstrated recombinational rescue of primer binding site (PBS)-impaired Akv murine leukemia virus-based vectors involving initial priming on endogenous viral sequences and template switching during cDNA synthesis to obtain PBS complementarity in second-strand transfer of reverse transcription (Mikkelsen et al., J. Virol. 70:1439-1447, 1996). By use of the same forced recombination system, we have now found recombinant proviruses of different structures, suggesting that PBS knockout vectors may be rescued through initial priming on endogenous virus RNA, read-through of the mutated PBS during minus-strand synthesis, and subsequent second-strand transfer mediated by the R-U5 complementarity of the plus strand and the extended minus-strand DNA acceptor template. Mechanisms for R-U5-mediated second-strand transfer and its possible role in retrovirus replication and evolution are discussed.

M3 - Journal article

C2 - 9499117

SN - 0022-538X

VL - 72

SP - 2519

EP - 2525

JO - Journal of Virology

JF - Journal of Virology

IS - 3

ER -