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AimB Is a Small Protein Regulator of Cell Size and MreB Assembly

Author(s): Werner, John N; Shi, Handuo; Hsin, Jen; Huang, Kerwyn Casey; Gitai, Zemer; et al

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Abstract: The MreB actin-like cytoskeleton assembles into dynamic polymers that coordinate cell shape in many bacteria. In contrast to most other cytoskeleton systems, few MreB-interacting proteins have been well characterized. Here, we identify a small protein from Caulobacter crescentus, an assembly inhibitor of MreB (AimB). AimB overexpression mimics inhibition of MreB polymerization, leading to increased cell width and MreB delocalization. Furthermore, aimB appears to be essential, and its depletion results in decreased cell width and increased resistance to A22, a small-molecule inhibitor of MreB assembly. Molecular dynamics simulations suggest that AimB binds MreB at its monomer-monomer protofilament interaction cleft and that this interaction is favored for C. crescentus MreB over Escherichia coli MreB because of a closer match in the degree of opening with AimB size, suggesting coevolution of AimB with MreB conformational dynamics in C. crescentus. We support this model through functional analysis of point mutants in both AimB and MreB, photo-cross-linking studies with site-specific unnatural amino acids, and species-specific activity of AimB. Together, our findings are consistent with AimB promoting MreB dynamics by inhibiting monomer-monomer assembly interactions, representing a new mechanism for regulating actin-like polymers and the first identification of a non-toxin MreB assembly inhibitor. Because AimB has only 104 amino acids and small proteins are often poorly characterized, our work suggests the possibility of more bacterial cytoskeletal regulators to be found in this class. Thus, like FtsZ and eukaryotic actin, MreB may have a rich repertoire of regulators to tune its precise assembly and dynamics.
Publication Date: 2020
Electronic Publication Date: 4-May-2020
Citation: Werner, John N, Shi, Handuo, Hsin, Jen, Huang, Kerwyn Casey, Gitai, Zemer, Klein, Eric A. (2020). AimB Is a Small Protein Regulator of Cell Size and MreB Assembly.. Biophysical journal, 119 (3), 593 - 604. doi:10.1016/j.bpj.2020.04.029
DOI: doi:10.1016/j.bpj.2020.04.029
ISSN: 0006-3495
EISSN: 1542-0086
Pages: 593 - 604
Language: eng
Type of Material: Journal Article
Journal/Proceeding Title: Biophysical Journal
Version: Author's manuscript

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