Identification of Important Amino Acid Residues That Modulate Binding of Escherichia coli GroEL to Its Various Cochaperones

Corresponding author: Costa Georgopoulos, Département de Biochimie Médicale, Centre Médical Universitaire, Université de Genève, 1, rue Michel Servet, 1211 Geneva 4, Switzerland., costa.georgopoulos{at}medecine.unige.ch (E-mail)

Communicating editor: A. L. SONENSHEIN

Genetic experiments have shown that the GroEL/GroES chaperone machine of Escherichia coli is absolutely essential, not only for bacterial growth but also for the propagation of many bacteriophages including . The virulent bacteriophages T4 and RB49 are independent of the host GroES function, because they encode their own cochaperone proteins, Gp31 and CocO, respectively. E. coli groEL44 mutant bacteria do not form colonies above 42° nor do they propagate bacteriophages , T4, or RB49. We found that the vast majority (40/46) of spontaneous groEL44 temperature-resistant colonies at 43° were due to the presence of an intragenic suppressor mutation. These suppressors define 21 different amino acid substitutions in GroEL, each affecting one of 13 different amino acid residues. All of these amino acid residues are located at or near the hinge, which regulates the large en bloc movements of the GroEL apical domain. All of these intragenic suppressors support bacteriophages , T4, and RB49 growth to various extents in the presence of the groEL44 allele. Since it is known that the GroEL44 mutant protein does not interact effectively with Gp31, the suppressor mutations should enhance cochaperone binding. Analogous intragenic suppressor studies were conducted with the groEL673 temperature-sensitive allele.

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