Structure of the Haemophilus influenzae uvr-1+ gene: homology with other uvrC-like genes and characterization of the Haemophilus influenzae uvr-1 and uvr-2 mutations.

Abstract

A 3.9 kb Haemophilus influenzae genomic DNA fragment was cloned in plasmid pUC9 that partially complemented the ultraviolet light sensitivity (UVs) of Escherichia coli uvrC mutant hosts. This fragment also complemented the UVs of H. influenzae uvr-1 (DB112) and uvr-2 (DB116) mutants. It genetically transformed the latter mutants to wild type UV resistance. The nucleotide (nt) sequence of this fragment revealed 3 open reading frames (ORFs). ORF2, now designated uvr-1+ (1746 nt), shows significant similarity in both the nt and amino acid (aa) composition to 7 complete proven or putative uvrC gene sequences. Computer analysis of the DNA sequence revealed several possible regulatory motifs 5' to uvr-1+, including a putative LexA-binding site as an inverted SOS box, located within the 3' region of ORF1, (extensive homology to the E. coli CMP-KDO synthetase gene), upstream of uvr-1+. Further computer analysis has also predicted that the four putative active site amino acids, located in the C-terminal half of each protein, are each situated in an area of secondary structure that are highly conserved.