Abstract
Single-molecular techniques have characterized dynamics of molecular motors such as flagellum in bacteria and myosin, kinesin, and dynein in eukaryotes. We can apply these techniques to a motility machine of archaea, namely, the archaellum, composed of a thin helical filament and a rotary motor. Although the size of the motor hinders the characterization of its motor function under a conventional optical microscope, fluorescence-labeling techniques allow us to visualize the architecture and function of the archaellar filaments in real time. Furthermore, a tiny polystyrene bead attached to the filament enables the visualization of motor rotation through the bead rotation and quantification of biophysical properties such as speed and torque produced by the rotary motor imbedded in the cell membrane. In this chapter, I describe the details of the above biophysical method based on an optical microscope.
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
References
Veigel C, Schmidt CF (2011) Moving into the cell: single-molecule studies of molecular motors in complex environments. Nat Rev Mol Cell Biol 12(3):163–176. https://doi.org/10.1038/nrm3062
Miyata M, Robinson RC, Uyeda TQP et al (2020) Tree of motility – a proposed history of motility systems in the tree of life. Genes Cells 25(1):6–21. https://doi.org/10.1111/gtc.12737
Kinosita Y, Nishizaka T (2018) Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum. Biophys Physicobiol 15:121–128. https://doi.org/10.2142/biophysico.15.0_121
Desmond E, Brochier-Armanet C, Gribaldo S (2007) Phylogenomics of the archaeal flagellum: rare horizontal gene transfer in a unique motility structure. BMC Evol Biol 7:106. https://doi.org/10.1186/1471-2148-7-106
Schlesner M, Miller A, Streif S et al (2009) Identification of Archaea-specific chemotaxis proteins which interact with the flagellar apparatus. BMC Microbiol 9:56. https://doi.org/10.1186/1471-2180-9-56
Banerjee A, Tsai CL, Chaudhury P et al (2015) FlaF is a β-sandwich protein that anchors the archaellum in the archaeal cell envelope by binding the S-layer protein. Structure 23(5):863–872. https://doi.org/10.1016/j.str.2015.03.001
Tsai CL, Tripp P, Sivabalasarma S et al (2020) The structure of the periplasmic FlaG-FlaF complex and its essential role for archaellar swimming motility. Nat Microbiol 5(1):216–225. https://doi.org/10.1038/s41564-019-0622-3
Chaudhury P, Neiner T, D’Imprima E et al (2016) The nucleotide-dependent interaction of FlaH and FlaI is essential for assembly and function of the archaellum motor. Mol Microbiol 99(4):674–685. https://doi.org/10.1111/mmi.13260
Chaudhury P, van der Does C, Albers SV (2018) Characterization of the ATPase FlaI of the motor complex of the Pyrococcus furiosus archaellum and its interactions between the ATP-binding protein FlaH. PeerJ 6:e4984. https://doi.org/10.7717/peerj.4984
Reindl S, Ghosh A, Williams GJ et al (2013) Insights into FlaI functions in archaeal motor assembly and motility from structures, conformations, and genetics. Mol Cell 49(6):1069–1082. https://doi.org/10.1016/j.molcel.2013.01.014
Kinosita Y, Uchida N, Nakane D et al (2016) Direct observation of rotation and steps of the archaellum in the swimming halophilic archaeon Halobacterium salinarum. Nat Microbiol 1(11):16148. https://doi.org/10.1038/nmicrobiol.2016.148
Iwata S, Kinosita Y, Uchida N et al (2019) Motor torque measurement of Halobacterium salinarum archaellar suggests a general model for ATP-driven rotary motors. Commun Biol 2:199. https://doi.org/10.1038/s42003-019-0422-6
Kinosita Y, Mikami N, Li Z et al (2020) Motile ghosts of the halophilic archaeon, Haloferax volcanii. Proc Natl Acad Sci U S A 117(43):26766–26772. https://doi.org/10.1073/pnas.2009814117
Kinosita Y, Ishida T, Yoshida M et al (2020) Distinct chemotactic behavior in the original Escherichia coli K-12 depending on forward-and-backward swimming, not on run-tumble movements. Sci Rep 10(1):15887. https://doi.org/10.1038/s41598-020-72429-1
Kinosita Y, Kikuchi Y, Mikami N et al (2018) Unforeseen swimming and gliding mode of an insect gut symbiont, Burkholderia sp. RPE64, with wrapping of the flagella around its cell body. ISME J 12(3):838–848. https://doi.org/10.1038/s41396-017-0010-z
Nishizaka T, Mizutani K, Masaike T (2007) Single-molecule observation of rotation of F1-ATPase through microbeads. Methods Mol Biol 392:171–181. https://doi.org/10.1007/978-1-59745-490-2_12
Li Z, Kinosita Y, Rodriguez-Franco M et al (2019) Positioning of the motility machinery in halophilic archaea. mBio 10:3. https://doi.org/10.1128/mBio.00377-19
Kohori A, Chiwata R, Hossain MD et al (2011) Torque generation in F1-ATPase devoid of the entire amino-terminal helix of the rotor that fills half of the stator orifice. Biophys J 101(1):188–195. https://doi.org/10.1016/j.bpj.2011.05.008
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kinosita, Y. (2023). Direct Observation of Archaellar Motor Rotation by Single-Molecular Imaging Techniques. In: Minamino, T., Miyata, M., Namba, K. (eds) Bacterial and Archaeal Motility. Methods in Molecular Biology, vol 2646. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3060-0_17
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3060-0_17
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3059-4
Online ISBN: 978-1-0716-3060-0
eBook Packages: Springer Protocols