Polymerization cycle of actin homolog MreB from a Gram-positive bacterium

Carballido-Lopez, R.; Chastanet, A.; Mao, W.; Renner, L. D.; Cornilleau, C.; Li de la Sierra-Gallay, I.; Benlamara, S.; Ah-Seng, Y.; Van Tilbeurgh, H.; Nessler, S.; Bertin, A.

doi:10.1101/2022.10.19.512861

bioRxiv · 10.1101/2022.10.19.512861

Polymerization cycle of actin homolog MreB from a Gram-positive bacterium

Carballido-Lopez, R.·Chastanet, A.·Mao, W.·Renner, L. D.·Cornilleau, C.·Li de la Sierra-Gallay, I.·Benlamara, S.·Ah-Seng, Y.·Van Tilbeurgh, H.·Nessler, S.·Bertin, A.

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Abstract

In most rod-shaped bacteria, the actin homologue MreB is an essential component of the protein complex effecting cell wall elongation. The polymerization cycle and filament properties of eukaryotic actin have studied for decades and are well characterized. However, purification and in vitro work on MreB proteins have proven very difficult. Current knowledge of MreB biochemical and polymerization properties remains limited and is based on MreB proteins from Gram-negative species. In this study, we report the first observation of organized filaments and the first 3D-structure of MreB from a Gram-positive bacterium. We have purified MreB from the thermophilic Geobacillus stearothermophilus and shown that it forms straight pairs of protofilaments in vitro, and that polymerization depends on the presence of both lipids and nucleotide triphosphate. Two spatially close short hydrophobic sequences mediate membrane anchoring. Importantly, we demonstrate that unlike eukaryotic actin, nucleotide hydrolysis is a prerequisite for MreB interaction with the membrane, and that binding to lipids then triggers polymerization. Based on our results, we propose a molecular model for the mechanism of MreB polymerization.

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Carballido-Lopez, R., Chastanet, A., Mao, W., Renner, L. D., Cornilleau, C., Li de la Sierra-Gallay, I., Benlamara, S., Ah-Seng, Y., Van Tilbeurgh, H., Nessler, S., Bertin, A.. 2022-10-20. Polymerization cycle of actin homolog MreB from a Gram-positive bacterium. https://doi.org/10.1101/2022.10.19.512861

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