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Chaperonin overexpression promotes genetic variation and enzyme evolution

Nature volume 459, pages 668–673 (2009)Cite this article

Abstract

Most protein mutations, and mutations that alter protein functions in particular, undermine stability and are therefore deleterious. Chaperones, or heat-shock proteins, are often implicated in buffering mutations, and could thus facilitate the acquisition of neutral genetic diversity and the rate of adaptation. We examined the ability of the Escherichia coli GroEL/GroES chaperonins to buffer destabilizing and adaptive mutations. Here we show that mutational drifts performed in vitro with four different enzymes indicated that GroEL/GroES overexpression doubled the number of accumulating mutations, and promoted the folding of enzyme variants carrying mutations in the protein core and/or mutations with higher destabilizing effects (destabilization energies of >3.5 kcal mol -1, on average, versus ∼1 kcal mol -1 in the absence of GroEL/GroES). The divergence of modified enzymatic specificity occurred much faster under GroEL/GroES overexpression, in terms of the number of adapted variants (≥2-fold) and their improved specificity and activity (≥10-fold). These results indicate that protein stability is a major constraint in protein evolution, and buffering mechanisms such as chaperonins are key in alleviating this constraint.

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Figure 1: The experimental setup.
Figure 2: GAPDH mutation-accumulation experiments.
Figure 3: Changes in activity and protein levels of enzyme variants expressed with and without GroEL/GroES overexpression.
Figure 4: Adaptive evolution of P. diminuta PTE towards higher esterase activity.

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Acknowledgements

Financial support by the EU via the MiFEM consortium, NIH grant number W81XWH-07-2-0020, and Israel Science Foundation are gratefully acknowledged. We are thankful to P. Goloubinoff for his advice, V. Kolotonouv for technical assistance, L. Serrano for the use of FoldX, and A. Horovitz and S. Rutherford for their help in refining this manuscript.

Author Contributions N.T. and D.S.T. designed the study, analysed the data and wrote the manuscript. N.T. performed the experiments.

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  1. Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel

    Nobuhiko Tokuriki & Dan S. Tawfik

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  1. Nobuhiko Tokuriki
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Correspondence to Dan S. Tawfik.

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Tokuriki, N., Tawfik, D. Chaperonin overexpression promotes genetic variation and enzyme evolution. Nature 459, 668–673 (2009). https://doi.org/10.1038/nature08009

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