Abstract
The stator is an eminent component of the flagellar motor and determines a number of the motor’s properties, such as the rotation-energizing coupling ion (H+ or Na+) or the torque that can be generated. The stator consists of several units located in the cytoplasmic membrane surrounding the flagellar drive shaft. Studies on flagellar motors of several bacterial species have provided evidence that the number as well as the retention time of stators coupled to the motor is highly dynamic and depends on the environmental conditions. Notably, numerous species possess more than a single distinct set of stators. It is likely that the presence of different stator units enables these bacteria to adjust the flagellar motor properties and function to meet the environmental requirements. One of these species is Shewanella oneidensis MR-1 that is equipped with a single polar flagellum and two stator units, the Na+-dependent PomAB and the H+-dependent MotAB. Here, we describe a method to determine stator dynamics by fluorescence microscopy, demonstrating how bacteria can change the composition of an intricate molecular machine according to environmental conditions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Minamino T, Imada K (2015) The bacterial flagellar motor and its structural diversity. Trends Microbiol 23:267–274
Minamino T, Imada K, Namba K (2008) Molecular motors of the bacterial flagella. Curr Opin Struct Biol 18:693–701
Sowa Y, Berry RM (2008) Bacterial flagellar motor. Q Rev Biophys 41:103–132
Stock D, Namba K, Lee LK (2012) Nanorotors and self-assembling macromolecular machines: the torque ring of the bacterial flagellar motor. Curr Opin Biotechnol 23:545–554
Beeby M, Ribardo DA, Brennan CA et al (2016) Diverse high-torque bacterial flagellar motors assemble wider stator rings using a conserved protein scaffold. Proc Natl Acad Sci U S A 113:1917–1926
Leake MC, Chandler JH, Wadhams GH et al (2006) Stoichiometry and turnover in single, functioning membrane protein complexes. Nature 443:355–358
Reid SW, Leake MC, Chandler JH et al (2006) The maximum number of torque-generating units in the flagellar motor of Escherichia coli is at least 11. Proc Natl Acad Sci U S A 103:8066–8071
Kojima S (2015) Dynamism and regulation of the stator, the energy conversion complex of the bacterial flagellar motor. Curr Opin Microbiol 28:66–71
Fung DC, Berg HC (1995) Powering the flagellar motor of Escherichia coli with an external voltage source. Nature 375:809–812
Sowa Y, Rowe AD, Leake MC et al (2005) Direct observation of steps in rotation of the bacterial flagellar motor. Nature 437:916–919
Tipping MJ, Steel BC, Delalez NJ et al (2013) Quantification of flagellar motor stator dynamics through in vivo proton-motive force control. Mol Microbiol 87:338–347
Tipping MJ, Delalez NJ, Lim R et al (2013) Load-dependent assembly of the bacterial flagellar motor. MBio 4:e00551-13. doi:10.1128/mBio.00551-13
Lele PP, Hosu BG, Berg HC (2013) Dynamics of mechanosensing in the bacterial flagellar motor. Proc Natl Acad Sci U S A 110:11839–11844
Fukuoka H, Wada T, Kojima S et al (2009) Sodium-dependent dynamic assembly of membrane complexes in sodium-driven flagellar motors. Mol Microbiol 71:825–835
Paulick A, Delalez NJ, Brenzinger S et al (2015) Dual stator dynamics in the Shewanella oneidensis MR-1 flagellar motor. Mol Microbiol 96:993–1001
Paulick A, Koerdt A, Lassak J et al (2009) Two different stator systems drive a single polar flagellum in Shewanella oneidensis MR-1. Mol Microbiol 71:836–850
Thormann KM, Paulick A (2010) Tuning the flagellar motor. Microbiology 156:1275–1283
Meijering E, Dzyubachyk O, Smal I (2012) Methods for cell and particle tracking. Methods Enzymol 504:183–200
Schindelin J, Arganda-Carreras I, Frise E et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682
Lassak J, Henche AL, Binnenkade L, Thormann KM (2010) ArcS, the cognate sensor kinase in an atypical Arc system of Shewanella oneidensis MR-1. Appl Environ Microbiol 76:3263–3274
Gibson DG, Young L, Chuang RY et al (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6:343–345
Brenzinger S, Dewenter L, Delalez NJ et al (2016) Mutations targeting the plug-domain of the Shewanella oneidensis proton-driven stator allow swimming at increased viscosity and under anaerobic conditions. Mol Microbiol 102:925–938
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Brenzinger, S., Thormann, K.M. (2017). Dynamics in the Dual Fuel Flagellar Motor of Shewanella oneidensis MR-1 . In: Minamino, T., Namba, K. (eds) The Bacterial Flagellum. Methods in Molecular Biology, vol 1593. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6927-2_23
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6927-2_23
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6926-5
Online ISBN: 978-1-4939-6927-2
eBook Packages: Springer Protocols