Abstract
With the rapid-freeze, deep-etch replica technique, the structural conformations of outer dynein arms in demembranated cilia from Tetrahymena were analyzed under two different conditions, i.e., in the absence of ATP and in the presence of ATP and vanadate. In the absence of ATP, the lateral view of axonemes was characterized by the egg- shaped outer dynein arms, which showed a slightly baseward tilt with a mean inclination of 11.1 degrees +/- 3.4 degrees SD from the perpendicular to the doublet microtubules. On the other hand, in the presence of 1 mM ATP and 100 microM vanadate, the outer arms were extended and slender and showed an increased baseward tilt with a mean inclination of 31.6 degrees +/- 4.9 degrees SD. In ATP-activated axonemes, these two types of arms coexisted, each type occurring in groups along one row of outer arms. These findings strongly suggest that the interdoublet sliding is caused by dynamic structural changes of dynein arms that follow the hydrolysis of ATP.
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- Berenblum I., Chain E. An improved method for the colorimetric determination of phosphate. Biochem J. 1938 Feb;32(2):295–298. doi: 10.1042/bj0320295. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibbons I. R. Chemical dissection of cilia. Arch Biol (Liege) 1965;76(2):317–352. [PubMed] [Google Scholar]
- Gibbons I. R. Cilia and flagella of eukaryotes. J Cell Biol. 1981 Dec;91(3 Pt 2):107s–124s. doi: 10.1083/jcb.91.3.107s. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibbons I. R., Cosson M. P., Evans J. A., Gibbons B. H., Houck B., Martinson K. H., Sale W. S., Tang W. J. Potent inhibition of dynein adenosinetriphosphatase and of the motility of cilia and sperm flagella by vanadate. Proc Natl Acad Sci U S A. 1978 May;75(5):2220–2224. doi: 10.1073/pnas.75.5.2220. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goodenough U. W., Heuser J. E. Substructure of the outer dynein arm. J Cell Biol. 1982 Dec;95(3):798–815. doi: 10.1083/jcb.95.3.798. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heuser J. E., Reese T. S., Dennis M. J., Jan Y., Jan L., Evans L. Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release. J Cell Biol. 1979 May;81(2):275–300. doi: 10.1083/jcb.81.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heuser J. E., Salpeter S. R. Organization of acetylcholine receptors in quick-frozen, deep-etched, and rotary-replicated Torpedo postsynaptic membrane. J Cell Biol. 1979 Jul;82(1):150–173. doi: 10.1083/jcb.82.1.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Okuno M. Inhibition and relaxation of sea urchin sperm flagella by vanadate. J Cell Biol. 1980 Jun;85(3):712–725. doi: 10.1083/jcb.85.3.712. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sale W. S., Gibbons I. R. Study of the mechanism of vanadate inhibition of the dynein cross-bridge cycle in sea urchin sperm flagella. J Cell Biol. 1979 Jul;82(1):291–298. doi: 10.1083/jcb.82.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sale W. S., Satir P. Direction of active sliding of microtubules in Tetrahymena cilia. Proc Natl Acad Sci U S A. 1977 May;74(5):2045–2049. doi: 10.1073/pnas.74.5.2045. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Satir P. Studies on cilia. 3. Further studies on the cilium tip and a "sliding filament" model of ciliary motility. J Cell Biol. 1968 Oct;39(1):77–94. doi: 10.1083/jcb.39.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Satir P., Wais-Steider J., Lebduska S., Nasr A., Avolio J. The mechanochemical cycle of the dynein arm. Cell Motil. 1981;1(3):303–327. doi: 10.1002/cm.970010304. [DOI] [PubMed] [Google Scholar]
- Summers K. E., Gibbons I. R. Adenosine triphosphate-induced sliding of tubules in trypsin-treated flagella of sea-urchin sperm. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3092–3096. doi: 10.1073/pnas.68.12.3092. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takahashi M., Tonomura Y. Binding of 30s dynein with the B-tubule of the outer doublet of axonemes from Tetrahymena pyriformis and adenosine triphosphate-induced dissociation of the complex. J Biochem. 1978 Dec;84(6):1339–1355. doi: 10.1093/oxfordjournals.jbchem.a132256. [DOI] [PubMed] [Google Scholar]
- Tsukita S., Tsukita S., Usukura J., Ishikawa H. Myosin filaments in smooth muscle cells of the guinea pig taenia coli: a freeze-substitution study. Eur J Cell Biol. 1982 Oct;28(2):195–201. [PubMed] [Google Scholar]
- Tsukita S., Usukura J., Tsukita S., Ishikawa H. The cytoskeleton in myelinated axons: a freeze-etch replica study. Neuroscience. 1982;7(9):2135–2147. doi: 10.1016/0306-4522(82)90125-7. [DOI] [PubMed] [Google Scholar]
- Warner F. D., Mitchell D. R., Perkins C. R. Structural conformation of the ciliary ATPase dynein. J Mol Biol. 1977 Aug 15;114(3):367–384. doi: 10.1016/0022-2836(77)90255-8. [DOI] [PubMed] [Google Scholar]
- Warner F. D., Mitchell D. R. Structural conformation of ciliary dynein arms and the generation of sliding forces in Tetrahymena cilia. J Cell Biol. 1978 Feb;76(2):261–277. doi: 10.1083/jcb.76.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]