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Nucleic Acids Research, 2000, Vol. 28, No. 3 791-799
© 2000 Oxford University Press

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

Anders H. Lund1, Mogens Duch1 and Finn Skou Pedersen1,2,*

1Department of Molecular and Structural Biology and 2Department of Medical Microbiology and Immunology, University of Aarhus, DK-8000, Denmark

Retroviral reverse transcription is initiated from a cellular tRNA molecule and all known exogenous isolates of murine leukemia virus utilise a tRNAPro molecule. While several studies suggest flexibility in murine leukemia virus primer utilisation, studies on human immunodeficiency virus and avian retro­viruses have revealed evidence of molecular adapt­ation 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 tRNAPro, we also retrieved PBS sequences matching four other tRNA molecules and demonstrate that Akv murine leukemia virus vectors may efficiently replicate using tRNAArg(CCU), tRNAPhe(GAA) and a hitherto unknown human tRNASer(CGA).

* To whom correspondence should be addressed at: Department of Molecular and Structural Biology, University of Aarhus, C.F. Møllers Allé, Building 130, DK-8000 Aarhus C, Denmark. Tel: +45 8942 2614; Fax: +45 8619 6500; Email: fsp@mbio.aau.dk Present address: Anders H. Lund, Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands


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