MUTATED PRIMER BINDING-SITES INTERACTING WITH DIFFERENT TRANSFER-RNASALLOW EFFICIENT MURINE LEUKEMIA-VIRUS REPLICATION

Two Akv murine leukemia virus-based retroviral vectors with primer bin ding sites matching tRNA(Gln-1) and tRNA(Lys-3) were constructed. The transduction efficiency of these mutated vectors was found to be compa rable to that of a vector carrying the wild-type primer binding site m atching tRNA(Pro). Polymerase chain reaction amplification and sequenc e analysis of transduced proviruses confirmed the transfer of vectors with mutated primer binding sites and further showed that tRNA(Gln-2) may act efficiently in conjunction with the tRNA(Gln-1) primer binding site. We conclude that murine leukemia virus can replicate by using v arious tRNA molecules as primers and propose primer binding site-tRNA primer interactions to be of major importance for tRNA primer selectio n. However, efficient primer selection does not require perfect Watson -Crick base pairing at all 18 positions of the primer binding site.