Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Jun 1;125(5):1109–1117. doi: 10.1083/jcb.125.5.1109

Mutations in the "dynein regulatory complex" alter the ATP-insensitive binding sites for inner arm dyneins in Chlamydomonas axonemes

PMCID: PMC2120054  PMID: 8195292

Abstract

To understand mechanisms of regulation of dynein activity along and around the axoneme we further characterized the "dynein regulatory complex" (drc). The lack of some axonemal proteins, which together are referred to as drc, causes the suppression of flagellar paralysis of radial spoke and central pair mutants. The drc is also an adapter involved in the ATP-insensitive binding of I2 and I3 inner dynein arms to doublet microtubules. Evidence supporting these conclusions was obtained through analyses of five drc mutants: pf2, pf3, suppf3, suppf4, and suppf5. Axonemes from drc mutants lack part of I2 and I3 inner dynein arms as well as subsets of seven drc components (apparent molecular weight from 29,000 to 192,000). In the absence of ATP-Mg, dynein-depleted axonemes from the same mutants bind I2 and I3 inner arms at both ATP-sensitive and -insensitive sites. At ATP-insensitive sites, they bind I2 and I3 inner arms to an extent that depends on the drc defect. This evidence suggested to us that the drc forms one binding site for the I2 and I3 inner arms on the A part of doublet microtubules.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams G. M., Huang B., Piperno G., Luck D. J. Central-pair microtubular complex of Chlamydomonas flagella: polypeptide composition as revealed by analysis of mutants. J Cell Biol. 1981 Oct;91(1):69–76. doi: 10.1083/jcb.91.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brokaw C. J., 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. [DOI] [PubMed] [Google Scholar]
  3. Brokaw C. J., Luck D. J., Huang B. Analysis of the movement of Chlamydomonas flagella:" the function of the radial-spoke system is revealed by comparison of wild-type and mutant flagella. J Cell Biol. 1982 Mar;92(3):722–732. doi: 10.1083/jcb.92.3.722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. EBERSOLD W. T., LEVINE R. P. A genetic analysis of linkage group I of Chlamydomonas reinhardi. Z Vererbungsl. 1959;90(1):74–82. doi: 10.1007/BF00888575. [DOI] [PubMed] [Google Scholar]
  5. Hosokawa Y., Miki-Noumura T. Bending motion of Chlamydomonas axonemes after extrusion of central-pair microtubules. J Cell Biol. 1987 Sep;105(3):1297–1301. doi: 10.1083/jcb.105.3.1297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Huang B., Piperno G., Ramanis Z., Luck D. J. Radial spokes of Chlamydomonas flagella: genetic analysis of assembly and function. J Cell Biol. 1981 Jan;88(1):80–88. doi: 10.1083/jcb.88.1.80. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Huang B., Ramanis Z., Luck D. J. Suppressor mutations in Chlamydomonas reveal a regulatory mechanism for Flagellar function. Cell. 1982 Jan;28(1):115–124. doi: 10.1016/0092-8674(82)90381-6. [DOI] [PubMed] [Google Scholar]
  8. Kurimoto E., Kamiya R. Microtubule sliding in flagellar axonemes of Chlamydomonas mutants missing inner- or outer-arm dynein: velocity measurements on new types of mutants by an improved method. Cell Motil Cytoskeleton. 1991;19(4):275–281. doi: 10.1002/cm.970190406. [DOI] [PubMed] [Google Scholar]
  9. Luck D., Piperno G., Ramanis Z., Huang B. Flagellar mutants of Chlamydomonas: studies of radial spoke-defective strains by dikaryon and revertant analysis. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3456–3460. doi: 10.1073/pnas.74.8.3456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mastronarde D. N., O'Toole E. T., McDonald K. L., McIntosh J. R., Porter M. E. Arrangement of inner dynein arms in wild-type and mutant flagella of Chlamydomonas. J Cell Biol. 1992 Sep;118(5):1145–1162. doi: 10.1083/jcb.118.5.1145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mitchell D. R., Rosenbaum J. L. A motile Chlamydomonas flagellar mutant that lacks outer dynein arms. J Cell Biol. 1985 Apr;100(4):1228–1234. doi: 10.1083/jcb.100.4.1228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Piperno G., Mead K., Shestak W. The inner dynein arms I2 interact with a "dynein regulatory complex" in Chlamydomonas flagella. J Cell Biol. 1992 Sep;118(6):1455–1463. doi: 10.1083/jcb.118.6.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Piperno G., Ramanis Z., Smith E. F., Sale W. S. Three distinct inner dynein arms in Chlamydomonas flagella: molecular composition and location in the axoneme. J Cell Biol. 1990 Feb;110(2):379–389. doi: 10.1083/jcb.110.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Piperno G., Ramanis Z. The proximal portion of Chlamydomonas flagella contains a distinct set of inner dynein arms. J Cell Biol. 1991 Feb;112(4):701–709. doi: 10.1083/jcb.112.4.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Porter M. E., Power J., Dutcher S. K. Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms. J Cell Biol. 1992 Sep;118(5):1163–1176. doi: 10.1083/jcb.118.5.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Smith E. F., Sale W. S. Structural and functional reconstitution of inner dynein arms in Chlamydomonas flagellar axonemes. J Cell Biol. 1992 May;117(3):573–581. doi: 10.1083/jcb.117.3.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Witman G. B. Isolation of Chlamydomonas flagella and flagellar axonemes. Methods Enzymol. 1986;134:280–290. doi: 10.1016/0076-6879(86)34096-5. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES