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Cited by 9Bacterial Genetics and BiotechnologyATP Synthase and ATPases ResearchProtein Structure and Dynamics

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Ancestral reconstruction of the MotA stator subunit reveals that conserved residues far from the pore are required to drive flagellar motility

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Islam, Md Imtiazul, Ridone, Pietro, Lin, Angela, Michie, Katharine A, Matzke, Nicholas J, Hochberg, Georg, Baker, Matthew A B·2023·microLife
DOI 10.1093/femsml/uqad011PMC PMC10117855

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Abstract

The bacterial flagellar motor (BFM) is a rotary nanomachine powered by the translocation of ions across the inner membrane through the stator complex. The stator complex consists of two membrane proteins: MotA and MotB (in H+-powered motors), or PomA and PomB (in Na+-powered motors). In this study, we used ancestral sequence reconstruction (ASR) to probe which residues of MotA correlate with function and may have been conserved to preserve motor function. We reconstructed 10 ancestral sequences of MotA and found four of them were motile in combination with contemporary Escherichia coli MotB and in combination with our previously published functional ancestral MotBs. Sequence comparison between wild-type (WT) E. coli MotA and MotA-ASRs revealed 30 critical residues across multiple domains of MotA that were conserved among all motile stator units. These conserved residues included pore-facing, cytoplasm-facing, and MotA–MotA intermolecular facing sites. Overall, this work demonstrates the role of ASR in assessing conserved variable residues in a subunit of a molecular complex.

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Metadata

Authors
Islam, M. I., Ridone, P., Lin, A., Michie, K. A., Matzke, N. J., Hochberg, G., & Baker, M. A. B.
Affiliations
  • UNSW Sydney (AU)
  • University of Auckland (NZ)
  • Philipps University of Marburg (DE)
Corresponding author
Baker, Matthew A B
ORCID
Venue
microLife
Publisher
Oxford University Press (OUP)
Published
2023
Volume
4
Pages
uqad011-uqad011
ISSN
2633-6693 (ISSN-L)
DOI
10.1093/femsml/uqad011
PMC
PMC10117855
OpenAlex
W4362568821
Cited by
9
References
1
Citations by year
2026: 1 · 2025: 2 · 2024: 3 · 2023: 3
FWCI
1.49
Type
journal article
Language
en
Open access
Licence
Locations
  • microLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • ResearchSpace (University of Auckland) (submittedVersion, cc-by)
Funders
  • Australian Research Council (190100165, DP190100497)
  • UNSW
  • CSIRO
  • Human Frontier Science Program
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
flagellar motor, MotB, ancestral sequence reconstruction, conserved residues, flagellar motility, Escherichia coli, ion translocation

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