title: 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.
abstract: 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)).
authors.affiliation: Department of Molecular and Structural Biology, University of Aarhus, Denmark.
authors.forename: A H
authors.initials: AH
authors.lastname: Lund
authors.forename: M
authors.initials: M
authors.lastname: Duch
authors.forename: F S
authors.initials: FS
authors.lastname: Pedersen
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D015982:Bias; D001665:Binding Sites; D015152:Blotting, Northern; D002460:Cell Line; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics* / Q000502:physiology; D051379:Mice; D009693:Nucleic Acid Hybridization / Q000235:genetics; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics*; D012343:RNA, Transfer / Q000235:genetics*; D012347:RNA, Transfer, Arg / Q000235:genetics; D012360:RNA, Transfer, Phe / Q000235:genetics; D012362:RNA, Transfer, Ser / Q000235:genetics; D011897:Random Allocation; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D013379:Substrate Specificity; D014779:Virus Replication / Q000235:genetics*
chemical_list: C034477:RNA primers; D012347:RNA, Transfer, Arg; D012360:RNA, Transfer, Phe; D012362:RNA, Transfer, Ser; D012313:RNA; D012343:RNA, Transfer