Genetic and molecular biological characterization of two homologous <italic>cheR</italic> genes from <italic>Leptospira interrogans</italic>

Abstract

The Leptospira interrogans genome encodes two copies of cheR genes and each of them is able to complement for the swarming defective phenotype of Escherichia coli cheR null mutant RP1254 to certain extent, while over-expression of either of them inhibits the swarming of the chemotactic wild-type E. coli strain, RP437. Therefore, both CheR1 and CheR2 ought to bear the methyltransferase activities, although CheR1 has only one instead of two conserved basic amino acid residues located on the positively charged face of α2-helix. When this residue as well as the Lys48 and Arg55 of CheR2 was mutated, none of the CheRs was able to maintain aforementioned complementation functions, suggesting their critical roles in recognition of methyl-accepting chemotaxis proteins similar to that of E. coli. Demonstrated by microarray assay, the expression of cheR1 in L. interrogans cultured at 28°C in Ellinghausen-McCullough-Johnson-Harris medium was significantly lower than the average transcription level of all other genes, while the transcription of cheR2 was significantly higher than that of cheR1 in accordance with real-time reverse transcriptase-polymerase chain reaction assay. Tandem MS-MS data mining for the proteome of the same culture detected 16 peptides derived from CheR2 but none from CheR1. Therefore, although both genes were shown to be functional in E. coli, the structurally more conserved CheR2 rather than CheR1 might be the major functional component of L. interrogans chemotaxis adaptation system under our laboratory culture conditions.