Phenotypic suppression methods for analyzing intra- and inter-molecular signaling interactions of chemoreceptors.

The receptors that mediate chemotactic behaviors in E. coli and other motile bacteria and archaea are exquisite molecular machines. They detect minute concentration changes in the organism's chemical environment, integrate multiple stimulus inputs, and generate a highly amplified output signal that modulates the cell's locomotor pattern. Genetic dissection and suppression analyses have played an important role in elucidating the molecular mechanisms that underlie chemoreceptor signaling. This chapter discusses three examples of phenotypic suppression analyses of receptor signaling defects. (i) Balancing suppression can occur in mutant receptors that have biased output signals and involves second-site mutations that create an offsetting bias change. Such suppressors can arise in many parts of the receptor and need not involve directly interacting parts of the molecule. (ii) Conformational suppression within a mutant receptor molecule occurs through a mutation that directly compensates for the initial structural defect. This form of suppression should be highly dependent on the nature of the structural alterations caused by the original mutation and its suppressor, but in practice may be difficult to distinguish from balancing suppression without high-resolution structural information about the mutant and pseudorevertant proteins. (iii) Conformational suppression between receptor molecules involves correction of a functional defect in one receptor by a mutational change in a heterologous receptor with which it normally interacts. The suppression patterns exhibit allele-specificity with respect to the compensatory residue positions and amino acid side chains, a hallmark of stereospecific protein-protein interactions.

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