Characterization of the cheY genes from Leptospira interrogans and their effects on the behavior of Escherichia coli.
The motility and chemotaxis system are critical for the virulence of pathogenic leptospire, which enable them to penetrate host tissue barriers during infection. The completed genome sequence of a representative virulent serovar type strain (Lai) of Leptospira interrogans serogroups Icterohaemorrhagiae (L. interrogans strain Lai) suggested that there were multiple copies of putative chemotaxis homologues located at its large chromosome. In order to verify the function of these proteins, the putative cheY genes were cloned into pQE31 vector and then expressed, respectively, in wild-type Escherichia coli strain RP437 and cheY defective strain RP5232. The results showed that all the five cheYs could restore the swarming of RP5232 strain to some extend. Overexpression of CheYs in RP437 showed inhibited swarming of RP437. To investigate the mechanism of chemotaxis signaling in L. interrogans strain Lai, certain aspartates (Asp-53, Asp-61, Asp-70, Asp-62, and Asp-66 for L. interrogans strain Lai CheY1, CheY2, CheY3, CheY4, and CheY5, respectively) were mutated. Expression of these mutated cheYs manifested neither restoration of the swarming ability of RP5232 nor inhibition on swarming ability of RP437. Multiple amino acid sequence alignment predicted ternary structures and the result of mutation experiment suggested that these conserved aspartate residues of L. interrogans were analogous to that in E. coli CheY in function and structure. So, L. interrogans and E. coli may have similar mechanisms of activation of the chemotaxis phosphorelay pathway, but there are differences in their control by signal terminator.
Li
Zhen-Hong
ZH
Department of Microbiology and Parasitology, 280 South Chongqing Road, Medical school, Shanghai Jiaotong University, Shanghai 200025, PR China.
Dong
Ke
K
Yuan
Jian-Ping
JP
Hu
Bao-Yu
BY
Liu
Jing-Xing
JX
Zhao
Guo-Ping
GP
Guo
Xiao-Kui
XK
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000494:pharmacology*; D001483:Base Sequence; D002633:Chemotaxis / Q000187:drug effects*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000187:drug effects* / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D007921:Leptospira interrogans / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects
D001426:Bacterial Proteins; D010750:Phosphoproteins; D001224:Aspartic Acid