Isolation and Characterization of the Enterobacter cloacae cheR Mutant Defective in Phosphate Taxis

Abstract

A chemotaxis-defective mutant of Enterobacter cloacae IFO3320, designated EC1, was isolated after N-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutagenesis. Computer-assisted capillary assays showed that EC1 failed to show chemotactic responses to peptone and inorganic phosphate (P_i). Cloning and sequence analysis showed that EC1 is a cheR mutant, suggesting that P_i taxis by E. cloacae is dependent on a methyl-accepting chemotaxis protein(s)(MCP). EC1 was further mutagenized with NTG to construct cheR pstS and cheR pstA double mutants. A recombinant plasmid pECT01.2, which contained the E. cloacae cheR gene, restored the ability of these double mutants to show chemotaxis toward peptone but not P_i. These results suggest that the phosphate-specific transport (Pst) system, together with a MCP(s), is required for detecting P_i in E. cloacae.