Selection of functional tRNA primers and primer binding site sequences from a retroviral combinatorial library: identification of new functional tRNA primers in murine leukemia virus replication.

TY - JOUR

T1 - Selection of functional tRNA primers and primer binding site sequences from a retroviral combinatorial library: identification of new functional tRNA primers in murine leukemia virus replication.

AU - Lund, Anders Henrik

AU - Duch, M

AU - Pedersen, F S

N1 - Keywords: 3T3 Cells; Animals; Base Sequence; Bias (Epidemiology); Binding Sites; Blotting, Northern; Cell Line; Gene Library; Genetic Vectors; Humans; Leukemia Virus, Murine; Mice; Nucleic Acid Hybridization; Polymerase Chain Reaction; RNA; RNA, Transfer; RNA, Transfer, Arg; RNA, Transfer, Phe; RNA, Transfer, Ser; Random Allocation; Regulatory Sequences, Nucleic Acid; Substrate Specificity; Virus Replication

PY - 2000

Y1 - 2000

N2 - Retroviral reverse transcription is initiated from a cellular tRNA molecule and all known exogenous isolates of murine leukemia virus utilise a tRNA(Pro)molecule. While several studies suggest flexibility in murine leukemia virus primer utilisation, studies on human immunodeficiency virus and avian retro-viruses have revealed evidence of molecular adapt-ation towards the specific tRNA isoacceptor used as replication primer. In this study, murine leukemia virus tRNA utilisation is investigated by in vivo screening of a retroviral vector combinatorial library with randomised primer binding sites. While most of the selected primer binding sites are complementary to the 3'-end of tRNA((Pro)), we also retrieved PBS sequences matching four other tRNA molecules and demonstrate that Akv murine leukemia virus vectors may efficiently replicate using tRNA(Arg(CCU)), tRNA(Phe(GAA))and a hitherto unknown human tRNA(Ser(CGA)).

AB - Retroviral reverse transcription is initiated from a cellular tRNA molecule and all known exogenous isolates of murine leukemia virus utilise a tRNA(Pro)molecule. While several studies suggest flexibility in murine leukemia virus primer utilisation, studies on human immunodeficiency virus and avian retro-viruses have revealed evidence of molecular adapt-ation towards the specific tRNA isoacceptor used as replication primer. In this study, murine leukemia virus tRNA utilisation is investigated by in vivo screening of a retroviral vector combinatorial library with randomised primer binding sites. While most of the selected primer binding sites are complementary to the 3'-end of tRNA((Pro)), we also retrieved PBS sequences matching four other tRNA molecules and demonstrate that Akv murine leukemia virus vectors may efficiently replicate using tRNA(Arg(CCU)), tRNA(Phe(GAA))and a hitherto unknown human tRNA(Ser(CGA)).

M3 - Journal article

C2 - 10637332

SN - 0305-1048

VL - 28

SP - 791

EP - 799

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 3

ER -