Crystal structures of CheY mutants Y106W and T871/Y106W - CheY activation correlates with movement of residue 106

Position 106 in CheY is highly conserved as an aromatic residue in the response regulator superfamily. In the structure of the wild-type, apo-CheY, Tyr(106) is a rotamer whose electron density is observed in both the inside and the outside positions. In the structure of the T87I mutant of CheY, the threonine to isoleucine change at position 87 causes the side chain of Tyr(106) to be elusively restricted to the outside position. In this report we demonstrate that the T87I mutation causes cells to be smooth swimming and non chemotactic. We also show that another CheY mutant, Y106W, causes cells to be more tumbly than wild-type CheY, and impairs chemotaxis. In the structure of Y106W, the side chain of Trp(106) stays exclusively in the inside position. Furthermore, a T87I/Y106W double mutant, which confers the same phenotype as T87I, restricts the side chain of Trp(106) to the outside position. The results from these behavioral and structural studies indicate that the rotameric nature of the Tyr(106) residue is involved in activation of the CheY molecule. Specifically, Chefs signaling ability correlates with the conformational heterogeneity of the Tyr(106) Side chain. Our data also suggest that these mutations affect the signal at an event subsequent to phosphorylation.