Sequence of the rec-2 locus of Haemophilus influenzae: homologies to comE-ORF3 of Bacillus subtilis and msbA of Escherichia coli

The nucleotide sequence of a 4243-bp PstI fragment containing the rec-2 gene of Haemophilus influenzae was determined. The amino acid (aa) sequences of four putative proteins were deduced from the corresponding open reading frames (ORFs). The 2400-bp ORF2 accounted for rec-2, based on the sequences of DNA fragments that contained rec-2::mini-Tn 10 mutations. The rec-2 gene encoded a putative 800-aa protein with a M r of 90561. Sequence analysis suggested that the rec-2 product contained nine highly probable integral membrane-spanning segments. Database searches showed that rec-2 was homologous to the comE-ORF3 gene of Bacillus subtilis. This hypothesis is consistent with the known involvement of both of these genes in the passage of transforming DNA through the competent-cell envelope. Although the sequences of the other three ORFs were incomplete, sufficient data were available to allow inferences about their homologies to other genes. ORF4, which overlapped ORF1, was homologous to the Escherichia coli dnaK suppresser gene, dksA, and therefore was named dsh-1 ( d naK s uppresser h omolog). Mutations in dsh-1 and its putative promoter region caused a mild sensitivity to UV light, but did not affect DNA recombination. ORF3, located downstream from rec-2, was homologous to msbA, an essential gene of E. coli with extensive similarity to the ATP-dependent translocators. ORF3 was named msh-1 ( ms bA h omolog). Mutations in msh-1 had no effects on genetic transformation. The close juxtaposition of rec-2 and msh-1 implied that the expression of msh-1 could be linked to the translation of the rec-2 ORF.