The fumarase genes of Escherichia coli: location of the fumB gene and discovery of a new gene (fumC)

Abstract

The fumB gene of Escherichia coli, which complements the fumarase deficiency of a fumA mutant when present in multiple copies, has been located at 93.5 min in the E. coli linkage map and its product has been identified as a polypeptide of 61 kDal. Four overlapping ColE1-fumB+ plasmids representing a continuous segment of 23.3 kb of bacterial DNA have been isolated from the Clarke-Carbon E. coli gene bank and the location of the fumB gene relative to the restriction map and the adjacent mel operon has been defined. Hybridization studies have shown that the fumB gene is homologous to the fumA gene, which complements the fumA1 mutation in single and multi-copy situations, and encodes an analogous 61 kDal product formerly regarded as the E. coli fumarase. The hybridization studies also showed that the Bacillus subtilis fumarase gene (citG) is homologous to an independent gene, fumC (formerly g48), which lies adjacent to the fumA gene at 35.5 min in the E. coli linkage map. The N-terminal sequences of the citG and fumC products exhibit a 51% identity over 88 residues. It is possible that the fumC and citG genes are fumarase structural genes of E. coli and B. subtilis, and that the fumA gene may encode a differentially-regulated fumarase or be a positive regulator gene which is essential for the expression of fumC (but not citG). If so, the fumB gene may encode a related enzyme or activator that can replace the fumA function when amplified.