Mechanical Properties of Inner-arm Dynein-f (dynein I1) Studied with in Vitro Motility Assays

Overview

Journal Biophys J

Publisher Cell Press

Specialty Biophysics

Date 2007 May 15

PMID 17496036

Citations 31

Authors

Norito Kotani

Hitoshi Sakakibara

Stan A Burgess

Hiroaki Kojima

Kazuhiro Oiwa

Affiliations

Soon will be listed here.

Abstract

Inner-arm dynein-f of Chlamydomonas flagella is a heterodimeric dynein. We performed conventional in vitro motility assays showing that dynein-f translocates microtubules at the comparatively low velocity of approximately 1.2 microm/s. From the dependence of velocity upon the surface density of dynein-f, we estimate its duty ratio to be 0.6-0.7. The relation between microtubule landing rate and surface density of dynein-f are well fitted by the first-power dependence, as expected for a processive motor. At low dynein densities, progressing microtubules rotate erratically about a fixed point on the surface, at which a single dynein-f molecule is presumably located. We conclude that dynein-f has high processivity. In an axoneme, however, slow and processive dynein-f could impede microtubule sliding driven by other fast dyneins (e.g., dynein-c). To obtain insight into the in vivo roles of dynein-f, we measured the sliding velocity of microtubules driven by a mixture of dyneins -c and -f at various mixing ratios. The velocity is modulated as a function of the ratio of dynein-f in the mixture. This modulation suggests that dynein-f acts as a load in the axoneme, but force pushing dynein-f molecules forward seems to accelerate their dissociation from microtubules.

Citing Articles

Structural diversity of axonemes across mammalian motile cilia.

Leung M, Sun C, Zeng J, Anderson J, Niu Q, Huang W Nature. 2025; 637(8048):1170-1177.

PMID: 39743588 PMC: 11779644. DOI: 10.1038/s41586-024-08337-5.

Active fluctuations of axoneme oscillations scale with number of dynein motors.

Sharma A, Friedrich B, Geyer V Proc Natl Acad Sci U S A. 2024; 121(46):e2406244121.

PMID: 39499635 PMC: 11573680. DOI: 10.1073/pnas.2406244121.

Versatile properties of dynein molecules underlying regulation in flagellar oscillation.

Fujiwara T, Shingyoji C, Higuchi H Sci Rep. 2023; 13(1):10514.

PMID: 37386019 PMC: 10310797. DOI: 10.1038/s41598-023-37242-6.

Axonemal structures reveal mechanoregulatory and disease mechanisms.

Walton T, Gui M, Velkova S, Fassad M, Hirst R, Haarman E Nature. 2023; 618(7965):625-633.

PMID: 37258679 PMC: 10266980. DOI: 10.1038/s41586-023-06140-2.

Composition and function of ciliary inner-dynein-arm subunits studied in Chlamydomonas reinhardtii.

Yamamoto R, Hwang J, Ishikawa T, Kon T, Sale W Cytoskeleton (Hoboken). 2021; 78(3):77-96.

PMID: 33876572 PMC: 8217317. DOI: 10.1002/cm.21662.

References

Smith E, Sale W . Microtubule binding and translocation by inner dynein arm subtype I1. Cell Motil Cytoskeleton. 1991; 18(4):258-68. DOI: 10.1002/cm.970180403. View

Sakakibara H, Kojima H, Sakai Y, Katayama E, Oiwa K . Inner-arm dynein c of Chlamydomonas flagella is a single-headed processive motor. Nature. 1999; 400(6744):586-90. DOI: 10.1038/23066. View

Pazour G, Witman G . The vertebrate primary cilium is a sensory organelle. Curr Opin Cell Biol. 2003; 15(1):105-10. DOI: 10.1016/s0955-0674(02)00012-1. View

Hancock W, Howard J . Processivity of the motor protein kinesin requires two heads. J Cell Biol. 1998; 140(6):1395-405. PMC: 2132675. DOI: 10.1083/jcb.140.6.1395. View

Burgess S, Walker M, Sakakibara H, Knight P, Oiwa K . Dynein structure and power stroke. Nature. 2003; 421(6924):715-8. DOI: 10.1038/nature01377. View

King S, Dutcher S . Phosphoregulation of an inner dynein arm complex in Chlamydomonas reinhardtii is altered in phototactic mutant strains. J Cell Biol. 1997; 136(1):177-91. PMC: 2132467. DOI: 10.1083/jcb.136.1.177. View

Vallee R . Reversible assembly purification of microtubules without assembly-promoting agents and further purification of tubulin, microtubule-associated proteins, and MAP fragments. Methods Enzymol. 1986; 134:89-104. DOI: 10.1016/0076-6879(86)34078-3. View

Cuda G, Pate E, Cooke R, Sellers J . In vitro actin filament sliding velocities produced by mixtures of different types of myosin. Biophys J. 1997; 72(4):1767-79. PMC: 1184371. DOI: 10.1016/S0006-3495(97)78823-4. View

Gibbons I . Cilia and flagella of eukaryotes. J Cell Biol. 1981; 91(3 Pt 2):107s-124s. PMC: 2112791. DOI: 10.1083/jcb.91.3.107s. View

10.

Webb M . A continuous spectrophotometric assay for inorganic phosphate and for measuring phosphate release kinetics in biological systems. Proc Natl Acad Sci U S A. 1992; 89(11):4884-7. PMC: 49192. DOI: 10.1073/pnas.89.11.4884. View

11.

Rikmenspoel R, Rudd W . The contractile mechanism in cilia. Biophys J. 1973; 13(9):955-93. PMC: 1484368. DOI: 10.1016/S0006-3495(73)86037-0. View

12.

Snell W, Pan J, Wang Q . Cilia and flagella revealed: from flagellar assembly in Chlamydomonas to human obesity disorders. Cell. 2004; 117(6):693-7. DOI: 10.1016/j.cell.2004.05.019. View

13.

Kojima H, Kikumoto M, Sakakibara H, Oiwa K . Mechanical Properties of a Single-Headed Processive Motor, Inner-Arm Dynein Subspecies-c of ChlamydomonasStudied at the Single Molecule Level. J Biol Phys. 2013; 28(3):335-45. PMC: 3456751. DOI: 10.1023/A:1020300319224. View

14.

Marshall W, Nonaka S . Cilia: tuning in to the cell's antenna. Curr Biol. 2006; 16(15):R604-14. DOI: 10.1016/j.cub.2006.07.012. View

15.

Shiroguchi K, Toyoshima Y . Regulation of monomeric dynein activity by ATP and ADP concentrations. Cell Motil Cytoskeleton. 2001; 49(4):189-99. DOI: 10.1002/cm.1032. View

16.

Brokaw C, Kamiya R . Bending patterns of Chlamydomonas flagella: IV. Mutants with defects in inner and outer dynein arms indicate differences in dynein arm function. Cell Motil Cytoskeleton. 1987; 8(1):68-75. DOI: 10.1002/cm.970080110. View

17.

Pazour G, Agrin N, Leszyk J, Witman G . Proteomic analysis of a eukaryotic cilium. J Cell Biol. 2005; 170(1):103-13. PMC: 2171396. DOI: 10.1083/jcb.200504008. View

18.

Howard J, Hudspeth A, Vale R . Movement of microtubules by single kinesin molecules. Nature. 1989; 342(6246):154-8. DOI: 10.1038/342154a0. View

19.

Burgess S, Walker M, Sakakibara H, Oiwa K, Knight P . The structure of dynein-c by negative stain electron microscopy. J Struct Biol. 2004; 146(1-2):205-16. DOI: 10.1016/j.jsb.2003.10.005. View

20.

Hirakawa E, Higuchi H, Toyoshima Y . Processive movement of single 22S dynein molecules occurs only at low ATP concentrations. Proc Natl Acad Sci U S A. 2000; 97(6):2533-7. PMC: 15963. DOI: 10.1073/pnas.050585297. View