Transcription of the group A streptococcal M12 protein gene (emm12) and the C5a peptidase gene (scpA), which encodes an inhibitor of complement-mediated chemotaxis, was previously shown to depend on a third genetic locus, designated virR. A 1.6 kb region of DNA which is 200 bp upstream of emm12 and is thought to contain the virR locus, was sequenced. An open reading frame which overlaps deletion mutations that define virR was identified. The sequence of the encoded VirR protein, which was deduced to contain 499 amino acids, is characteristic of cytoplasmic proteins. Comparison of the VirR protein to a variety of DNA binding proteins, such as lambda Cro, revealed a DNA binding motif. VirR was also compared to the M6 positive regulator, mry, and found to be 98% homologous. The predicted virR promoter is preceded by two sets of inverted repeats, in contrast to mry which is preceded by one repeat. Introduction of virR on the shuttle vector pAM401 into a strain of group A Streptococcus with a deletion in the chromosomal virR gene demonstrated that the VirR protein activated transcription of both emm12 and scpA genes in trans. Analysis of RNA by Northern blot using virR-specific probes identified two virR transcripts, a 1.6 kb transcript which corresponds to the predicted size of the gene, and a second transcript, 3.5 kb, which also overlaps virR. These results demonstrate that virR and mry are structurally and functionally very similar and show that the former is a trans activator of both M protein and C5a peptidase synthesis.