PAS/poly-HAMP signalling in Aer-2, a soluble haem-based sensor.
Poly-HAMP domains are widespread in bacterial chemoreceptors, but previous studies have focused on receptors with single HAMP domains. The Pseudomonas aeruginosa chemoreceptor, Aer-2, has an unusual domain architecture consisting of a PAS-sensing domain sandwiched between three N-terminal and two C-terminal HAMP domains, followed by a conserved kinase control module. The structure of the N-terminal HAMP domains was recently solved, making Aer-2 the first protein with resolved poly-HAMP structure. The role of Aer-2 in P. aeruginosa is unclear, but here we show that Aer-2 can interact with the chemotaxis system of Escherichia coli to mediate repellent responses to oxygen, carbon monoxide and nitric oxide. Using this model system to investigate signalling and poly-HAMP function, we determined that the Aer-2 PAS domain binds penta-co-ordinated b-type haem and that reversible signalling requires four of the five HAMP domains. Deleting HAMP 2 and/or 3 resulted in a kinase-off phenotype, whereas deleting HAMP 4 and/or 5 resulted in a kinase-on phenotype. Overall, these data support a model in which ligand-bound Aer-2 PAS and HAMP 2 and 3 act together to relieve inhibition of the kinase control module by HAMP 4 and 5, resulting in the kinase-on state of the Aer-2 receptor.
Division of Microbiology and Molecular Genetics, Loma Linda University, Loma Linda, CA 92350, USA. kwatts@llu.edu
Kylie J
KJ
Watts
Barry L
BL
Taylor
Mark S
MS
Johnson
D000222:Adaptation, Physiological / Q000187:drug effects; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D002248:Carbon Monoxide / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D006418:Heme / Q000378:metabolism*; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D015398:Signal Transduction / Q000187:drug effects; D012995:Solubility / Q000187:drug effects
C492263:ARR-2 protein, Pseudomonas aeruginosa; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D009569:Nitric Oxide; D006418:Heme; D002248:Carbon Monoxide; D010100:Oxygen
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