dc.contributor.author |
Ishida, Tsubasa |
en |
dc.contributor.author |
Ito, Rie |
en |
dc.contributor.author |
Clark, Jessica |
en |
dc.contributor.author |
Matzke, Nicholas |
en |
dc.contributor.author |
Sowa, Yoshiyuki |
en |
dc.contributor.author |
Baker, Matthew AB |
en |
dc.date.accessioned |
2019-09-29T21:58:37Z |
en |
dc.date.issued |
2019-06 |
en |
dc.identifier.issn |
0950-382X |
en |
dc.identifier.uri |
http://hdl.handle.net/2292/47990 |
en |
dc.description.abstract |
The bacterial flagellar motor powers the rotation that propels the swimming bacteria. Rotational torque is generated by harnessing the flow of ions through ion channels known as stators which couple the energy from the ion gradient across the inner membrane to rotation of the rotor. Here, we used error-prone PCR to introduce single point mutations into the sodium-powered Vibrio alginolyticus/Escherichia coli chimeric stator PotB and selected for motors that exhibited motility in the presence of the sodium-channel inhibitor phenamil. We found single mutations that enable motility under phenamil occurred at two sites: (i) the transmembrane domain of PotB, corresponding to the TM region of the PomB stator from V. alginolyticus and (ii) near the peptidoglycan binding region that corresponds to the C-terminal region of the MotB stator from E. coli. Single cell rotation assays confirmed that individual flagellar motors could rotate in up to 100 µM phenamil. Using phylogenetic logistic regression, we found correlation between natural residue variation and ion source at positions corresponding to PotB F22Y, but not at other sites. Our results demonstrate that it is not only the pore region of the stator that moderates motility in the presence of ion-channel blockers. |
en |
dc.format.medium |
Print-Electronic |
en |
dc.language |
eng |
en |
dc.relation.ispartofseries |
Molecular microbiology |
en |
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
en |
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
en |
dc.subject |
Flagella |
en |
dc.subject |
Escherichia coli |
en |
dc.subject |
Vibrio alginolyticus |
en |
dc.subject |
Sodium |
en |
dc.subject |
Amiloride |
en |
dc.subject |
Peptidoglycan |
en |
dc.subject |
Bacterial Proteins |
en |
dc.subject |
Escherichia coli Proteins |
en |
dc.subject |
ATP-Binding Cassette Transporters |
en |
dc.subject |
Sodium Channels |
en |
dc.subject |
Sodium Channel Blockers |
en |
dc.subject |
Phylogeny |
en |
dc.subject |
Point Mutation |
en |
dc.subject |
Torque |
en |
dc.subject |
Molecular Motor Proteins |
en |
dc.title |
Sodium-powered stators of the bacterial flagellar motor can generate torque in the presence of phenamil with mutations near the peptidoglycan-binding region. |
en |
dc.type |
Journal Article |
en |
dc.identifier.doi |
10.1111/mmi.14246 |
en |
pubs.issue |
6 |
en |
pubs.begin-page |
1689 |
en |
pubs.volume |
111 |
en |
dc.rights.holder |
Copyright: The author |
en |
pubs.end-page |
1699 |
en |
pubs.publication-status |
Published |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/RestrictedAccess |
en |
pubs.subtype |
Research Support, Non-U.S. Gov't |
en |
pubs.subtype |
Journal Article |
en |
pubs.elements-id |
770450 |
en |
pubs.org-id |
Science |
en |
pubs.org-id |
Biological Sciences |
en |
dc.identifier.eissn |
1365-2958 |
en |
pubs.record-created-at-source-date |
2019-03-31 |
en |
pubs.dimensions-id |
30927553 |
en |