CLONING AND SEQUENCING OF A 29-KB REGION OF THE BACILLUS-SUBTILIS GENOME CONTAINING THE HUT AND WAPA LOCI

Within the framework of an international project for the sequencing of the entire Bacillus subtilis genome, a 29 kb chromosome segment, whic h contains the hut operon (335 degrees) and the wapA gene, has been cl oned and sequenced, This region (28954 bp) contains 21 complete ORFs a nd one partial one. The 5th, 6th and 17th genes correspond to hutH enc oding histidase, hutP encoding the positive regulator for the hut oper on and wapA encoding a precursor of three major wall-associated protei ns, respectively. A homology search for their products deduced from th e 21 complete ORFs revealed that nine of them exhibit significant homo logy to known proteins such as urocanase (Pseudomonas putida), a prote in involved in clavulanic acid biosynthesis (Streptomyces griseus), am ino acid permeases (lysine, Escherichia coli; histidine, Saccharomyces cerevisiae; and others), beta-glucoside-specific phosphotransferases (E. coli and Erwinia chrysanthemi) and 6-phospho-beta-glucosidases (E. coli and Erw. chrysanthemi). Based on the features of the determined sequence and the results of the homology search, as well as on genetic data and sequence of the hut genes reported by other groups, it is pr edicted that the B. subtilis hut operon may consist of the following s ix genes (6th-1st), the last of which is followed by a typical p-indep endent transcription terminator: hutP, hutH, EE57A (hutU) encoding uro canase, EE57B (hutI) encoding imidazolone-5-propionate hydrolase, EE57 C (hutG) encoding formiminoglutamate hydrolase and EE57D (tentatively designated as hutM) possibly encoding histidine permease. Interestingl y, the direction of transcription of these hut genes is opposite to th at of the movement of the replication fork.