Mutations targeting the plug-domain of the Shewanella oneidensis proton-driven stator allow swimming at increased viscosity and under anaerobic conditions.
Shewanella oneidensis MR-1 possesses two different stator units to drive flagellar rotation, the Na+-dependent PomAB stator and the H+-driven MotAB stator, the latter possibly acquired by lateral gene transfer. Although either stator can independently drive swimming through liquid, MotAB-driven motors cannot support efficient motility in structured environments or swimming under anaerobic conditions. Using ΔpomAB cells we isolated spontaneous mutants able to move through soft agar. We show that a mutation that alters the structure of the plug domain in MotB affects motor functions and allows cells to swim through media of increased viscosity and under anaerobic conditions. The number and exchange rates of the mutant stator around the rotor were not significantly different from wild-type stators, suggesting that the number of stators engaged is not the cause of increased swimming efficiency. The swimming speeds of planktonic mutant MotAB-driven cells was reduced, and overexpression of some of these stators caused reduced growth rates, implying that mutant stators not engaged with the rotor allow some proton leakage. The results suggest that the mutations in the MotB plug domain alter the proton interactions with the stator ion channel in a way that both increases torque output and allows swimming at decreased pmf values.
Department of Microbiology and Molecular Biology at the IFZ, Justus-Liebig-Universität Gießen, Gießen, 35392, Germany.|Department of Ecophysiology, Max-Planck-Institut für terrestrische Mikrobiologie, Marburg, 35043, Germany.
Susanne
S
Brenzinger
Department of Physics, Universität Köln, Cologne, 50674, Germany.
Lena
L
Dewenter
Biochemistry Department, University of Oxford, Oxford, OX1 3QU, UK.
Nicolas J
NJ
Delalez
Philipps-Universität, Marburg, Germany LOEWE Center for Synthetic Microbiology, Marburg, 35043, Germany.
Oliver
O
Leicht
Department of Ecophysiology, Max-Planck-Institut für terrestrische Mikrobiologie, Marburg, 35043, Germany.
Volker
V
Berndt
Department of Ecophysiology, Max-Planck-Institut für terrestrische Mikrobiologie, Marburg, 35043, Germany.
Anja
A
Paulick
Physics Department, University of Oxford, Oxford, OX1 3QU, UK.
Richard M
RM
Berry
Philipps-Universität, Marburg, Germany LOEWE Center for Synthetic Microbiology, Marburg, 35043, Germany.|Max-Planck-Institut für terrestrische Mikrobiologie & LOEWE Center für Synthetische Mikrobiologie, Marburg, 35043, Germany.
Martin
M
Thanbichler
Biochemistry Department, University of Oxford, Oxford, OX1 3QU, UK.
Judith P
JP
Armitage
Department of Physics, Universität Köln, Cologne, 50674, Germany.
Berenike
B
Maier
Department of Microbiology and Molecular Biology at the IFZ, Justus-Liebig-Universität Gießen, Gießen, 35392, Germany.
Kai M
KM
Thormann
D000693:Anaerobiosis; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D005407:Flagella / Q000235:genetics* / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D011522:Protons; D020592:Shewanella / Q000235:genetics* / Q000378:metabolism; D014783:Viscosity
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D020409:Molecular Motor Proteins; C063010:MotA protein, Bacteria; C064779:MotB protein, Bacteria; C103352:PomA protein, bacteria; C103353:PomB protein, bacteria; D011522:Protons