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A counter-enzyme complex regulates glutamate metabolism in Bacillus subtilis

View ORCID ProfileVijay Jayaraman, D. John Lee, View ORCID ProfileNadav Elad, Shay Vimer, View ORCID ProfileMichal Sharon, View ORCID ProfileJames S. Fraser, View ORCID ProfileDan S. Tawfik
doi: https://doi.org/10.1101/2021.04.12.439528
Vijay Jayaraman
1Department of Biomolecular sciences, Weizmann Institute of Science, Rehovot, Israel, 7610001
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D. John Lee
3Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94158, USA
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Nadav Elad
2Department of chemical research support, Weizmann institute of Science, Rehovot, Israel, 7610001
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Shay Vimer
1Department of Biomolecular sciences, Weizmann Institute of Science, Rehovot, Israel, 7610001
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Michal Sharon
1Department of Biomolecular sciences, Weizmann Institute of Science, Rehovot, Israel, 7610001
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James S. Fraser
3Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California 94158, USA
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Dan S. Tawfik
1Department of Biomolecular sciences, Weizmann Institute of Science, Rehovot, Israel, 7610001
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  • For correspondence: dan.tawfik@weizmann.ac.il
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Summary

Multi-enzyme assemblies composed of metabolic enzymes catalyzing sequential reactions are being increasingly studied. Here, we report the discovery of a 1.6 megadalton multi-enzyme complex from Bacillus subtilis composed of two enzymes catalyzing opposite rather than sequential reactions (“counter-enzymes”): glutamate synthase (GltAB), and glutamate dehydrogenase (GudB), that make and break glutamate, respectively. In vivo and in vitro studies show that the primary role of complex formation is to inhibit GudB’s activity as this enzyme is constitutively expressed including in glutamate-limiting conditions. Using cryo-electron microscopy, we elucidated the structure of the complex and the basis of GudB’s inhibition. Finally, we show that this complex that exhibits unusual oscillatory progress curves is a necessity for planktonic growth in glutamate-limiting conditions, but is also essential for biofilm growth in glutamate-rich media, suggesting a regulatory role at fluctuating glutamate concentrations.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 12, 2021.
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A counter-enzyme complex regulates glutamate metabolism in Bacillus subtilis
Vijay Jayaraman, D. John Lee, Nadav Elad, Shay Vimer, Michal Sharon, James S. Fraser, Dan S. Tawfik
bioRxiv 2021.04.12.439528; doi: https://doi.org/10.1101/2021.04.12.439528
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A counter-enzyme complex regulates glutamate metabolism in Bacillus subtilis
Vijay Jayaraman, D. John Lee, Nadav Elad, Shay Vimer, Michal Sharon, James S. Fraser, Dan S. Tawfik
bioRxiv 2021.04.12.439528; doi: https://doi.org/10.1101/2021.04.12.439528

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