The Streptococcus gordonii (Challis) glucosyltransferase-encoding determinant gtfG is regulated by the product of the adjacent gene rgg. Results of analyses described here showed that in both S. gordonii and Escherichia coli Rgg is a positive transcriptional regulator of glucosyltransferase expression. In addition, the transcriptional start sites of both gtfG and rgg were determined.Glucosyltransferases (GTFs) are enzymes that polymerize the glucose moiety of sucrose to yield glucan. The enzymes are thought to be important for the mutans streptococci in sucrosedependent bacterial accumulation on smooth tooth surfaces (9). The production of glucan by Streptococcus gordonii has been correlated with the accumulation of this organism on hydroxyapatite beads (24); thus, GTF may play a role in the colonization of S. gordonii on the tooth surface. Experimental evidence also suggests that this enzyme may be a virulence factor for S. gordonii in the pathogenesis of infective endocarditis (12, 16), although a recent report suggests that this is not the case for S. gordonii Challis, at least in a rat endocarditis model (26).The complete nucleotide sequence of S. gordonii Challis gtfG was determined recently (25), and the corresponding protein had a mass of ca. 170 kDa. We previously identified a gene, rgg, located immediately upstream of gtfG, that functions as a positive regulator of GTF expression (21). Haisman and Jenkinson have also identified two distinct genetic regulatory loci, gtf-20 and gtf-30, that appear to regulate GTF expression (4). Here, we report the results of Northern (RNA) blot and primer extension analyses of the rgg-gtfG locus, which show (i) transcripts that are expressed at the rgg and gtfG loci and where they are initiated and (ii) that the rgg product affects GTF expression at the level of transcription. In addition, our conclusions are corroborated by the demonstration of regulation of gtfG in Escherichia coli with a gtfGЈ-lacZ transcriptional reporter fusion.A restriction endonuclease map of the rgg-gtfG region of the S. gordonii Challis chromosome, related probes used for Northern hybridization, and sizes and locations of transcripts detected are shown in Fig. 1A. Several mRNA transcripts were identified in the strain containing the streptococcal cloning vector pVA749 (Fig. 1B, lanes 4). A 1.7-kb band was identified with the upstream probe. A 1.1-kb band was identified exclusively with the rgg probe, a 5.4-kb band only was identified with the gtfG probe, and a 6.4-kb band was identified with both the rgg and gtfG probes. The data are consistent with the following three transcripts derived from the rgg and gtfG loci (Fig. 1A): (i) a 1.1-kb rgg transcript, large enough to contain the rgg open reading frame (ORF) of 891 nucleotides; (ii) a 5.4-kb gtfG transcript that is of a size sufficient to contain the entire structural determinant of the GTF (22, 25); and (iii) a 6.4-kb rgggtfG transcript that likely represents a transcript originating at the rgg promoter and terminating at the same site as the...