Identification and characterization of the aspartate chemosensory receptor of Campylobacter jejuni.
Campylobacter jejuni is a highly motile bacterium that responds via chemotaxis to environmental stimuli to migrate towards favourable conditions. Previous in silico analysis of the C. jejuni strain NCTC11168 genome sequence identified 10 open reading frames, tlp1-10, that encode putative chemosensory receptors. We describe the characterization of the role and specificity of the Tlp1 chemoreceptor (Cj1506c). In vitro and in vivo models were used to determine if Tlp1 had a role in host colonization. The tlp1(-) isogenic mutant was more adherent in cell culture, however, showed reduced colonization ability in chickens. Specific interactions between the purified sensory domain of Tlp1 and l-aspartate were identified using an amino acid array and saturation transfer difference nuclear magnetic resonance spectroscopy. Chemotaxis assays showed differences between migration of wild-type C. jejuni cells and that of a tlp1(-) isogenic mutant, specifically towards aspartate. Furthermore, using yeast two-hybrid and three-hybrid systems for analysis of protein-protein interactions, the cytoplasmic signalling domain of Tlp1 was found to preferentially interact with CheV, rather than the CheW homologue of the chemotaxis signalling pathway; this interaction was confirmed using immune precipitation assays. This is the first identification of an aspartate receptor in bacteria other than Escherichia coli and Salmonella enterica serovar Typhimurium.
Hartley-Tassell
Lauren E
LE
Institute for Glycomics, Griffith University, Gold Coast Campus, Australia.
Shewell
Lucy K
LK
Day
Christopher J
CJ
Wilson
Jennifer C
JC
Sandhu
Randeep
R
Ketley
Julian M
JM
Korolik
Victoria
V
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016123:Campylobacter jejuni / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002633:Chemotaxis / Q000235:genetics; D002645:Chickens / Q000382:microbiology; D008024:Ligands; D017434:Protein Structure, Tertiary; D017459:Receptors, Amino Acid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D008024:Ligands; D017459:Receptors, Amino Acid; C031959:aspartic acid receptor; D001224:Aspartic Acid