A stutter in the coiled coil domain of E.coli co-chaperone GrpE connects structure with function

Upadhyay, T.; Karekar, V. V.; Saraogi, I.

doi:10.1101/2020.07.03.186056

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bioRxiv · 10.1101/2020.07.03.186056

A stutter in the coiled coil domain of E.coli co-chaperone GrpE connects structure with function

Upadhyay, T.·Karekar, V. V.·Saraogi, I.

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Abstract

In bacteria, the co-chaperone GrpE acts as a nucleotide exchange factor and plays an important role in controlling the chaperone cycle of DnaK. The functional form of GrpE is an asymmetric dimer, consisting of a long non-ideal coiled-coil. During heat stress, this region partially unfolds and prevents DnaK nucleotide exchange, ultimately ceasing the chaperone cycle. In this study, we elucidate the role of thermal unfolding of the coiled-coil domain of E. coli GrpE in regulating its co-chaperonic activity. The presence of a stutter disrupts the regular heptad arrangement typically found in an ideal coiled coil resulting in structural distortion. Introduction of hydrophobic residues at the stutter altered the structural stability of the coiled-coil. Using an in vitro FRET assay, we show for the first time that the enhanced stability of GrpE resulted in an increased affinity for DnaK. However, the mutants were defective in in vitro functional assays, and were unable to support bacterial growth at heat shock temperature in a grpE-deleted E. coli strain. This work provides valuable insights into the functional role of a stutter in the GrpE coiled-coil, and its role in regulating the DnaK-chaperone cycle for bacterial survival during heat stress. More generally, our findings illustrate how a sequence specific stutter in a coiled-coil domain regulates the structure function trade-off in proteins.Competing Interest StatementThe authors have declared no competing interest.View Full Text

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Upadhyay, T., Karekar, V. V., Saraogi, I.. 2020-07-03. A stutter in the coiled coil domain of E.coli co-chaperone GrpE connects structure with function. https://doi.org/10.1101/2020.07.03.186056

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