Structure and biological activity of the subgenomic Mtv-6 endogenous provirus

Abstract

The Mtv-6 provirus has an incomplete genome, but retains a functional superantigen gene (sag) which directs the thymicdeletion of CD4 + T cells expressing T cell receptors containing the V β3 or V β5 chains. To better understand the Mtv-6 superantigen, the structure and biological activity of the Mtv-6 provirus was analyzed. First, the complete nucleotide sequence was determined, and the mutation producing the subgenomic provirus was identified. Second, the nucleotide sequence of the 5′ end of the sag gene transcript (including the splice junction) was determined by sequence analysis of a cDNA clone. Third, the superantigen activity of Mtv-6 was analyzed in mice carrying the Mtv-6 provirus isolated by selective breeding on a genetic background free of endogenous and exogenous mouse mammary tumor virus (MMTV). These studies demonstrate that (i) the Mtv-6 provirus contains a 6.2-kb deletion between two 12-bp direct repeats encompassing the central portion of the provirus but not affecting sag gene splicing or translation, (ii) the sag gene transcript has the structure predicted from previous S1 nuclease mapping studies, and (iii) the Mtv-6 superantigen can direct thymic deletion of target V β3 + and V β5 +T cells in the absence of gene products from full-length MMTV proviruses.