Home LiteratureArticle Details
PMID: 9843573 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The Mr 140,000 intermediate chain of Chlamydomonas flagellar inner arm dynein is a WD-repeat protein implicated in dynein arm anchoring.

Molecular biology of the cell ·Vol. 9 ·No. 12 ·1998-12-00 ·Pages 3335-49

Yang P, Sale WS

Abstract

Previous structural and biochemical studies have revealed that the inner arm dynein I1 is targeted and anchored to a unique site located proximal to the first radial spoke in each 96-nm axoneme repeat on flagellar doublet microtubules. To determine whether intermediate chains mediate the positioning and docking of dynein complexes, we cloned and characterized the 140-kDa intermediate chain (IC140) of the I1 complex. Sequence and secondary structural analysis, with particular emphasis on beta-sheet organization, predicted that IC140 contains seven WD repeats. Reexamination of other members of the dynein intermediate chain family of WD proteins indicated that these polypeptides also bear seven WD/beta-sheet repeats arranged in the same pattern along each intermediate chain protein. A polyclonal antibody was raised against a 53-kDa fusion protein derived from the C-terminal third of IC140. The antibody is highly specific for IC140 and does not bind to other dynein intermediate chains or proteins in Chlamydomonas flagella. Immunofluorescent microscopy of Chlamydomonas cells confirmed that IC140 is distributed along the length of both flagellar axonemes. In vitro reconstitution experiments demonstrated that the 53-kDa C-terminal fusion protein binds specifically to axonemes lacking the I1 complex. Chemical cross-linking indicated that IC140 is closely associated with a second intermediate chain in the I1 complex. These data suggest that IC140 contains domains responsible for the assembly and docking of the I1 complex to the doublet microtubule cargo.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Chlamydomonas/enzymology,genetics Cloning, Molecular DNA Primers/genetics DNA, Protozoan/genetics Dyneins/chemistry,genetics,metabolism Flagella/enzymology Genes, Protozoan Molecular Sequence Data Molecular Weight Recombinant Fusion Proteins/chemistry,genetics,metabolism Repetitive Sequences, Amino Acid Sequence Homology, Amino Acid
Chemicals
DNA Primers DNA, Protozoan Recombinant Fusion Proteins Dyneins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Yang P
Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Sale W S
References (58)
58 references, click to expand
  1. Polarity of flagellar assembly in Chlamydomonas.
    J Cell Biol. 1992 Dec;119(6):1605-11 PMID: 1281816
  2. PF20 gene product contains WD repeats and localizes to the intermicrotubule bridges in Chlamydomonas flagella.
    Mol Biol Cell. 1997 Mar;8(3):455-67 PMID: 9188098
  3. Isolation of the Chlamydomonas regulatory gene NIT2 by transposon tagging.
    Genetics. 1993 Jul;134(3):737-47 PMID: 8394263
  4. Assembly of flagellar radial spoke proteins in Chlamydomonas: identification of the axoneme binding domain of radial spoke protein 3.
    J Cell Biol. 1993 Oct;123(1):183-90 PMID: 8408197
  5. Molecular analysis of the gamma heavy chain of Chlamydomonas flagellar outer-arm dynein.
    J Cell Sci. 1994 Mar;107 ( Pt 3):497-506 PMID: 7516341
  6. Sequence analysis of the Chlamydomonas alpha and beta dynein heavy chain genes.
    J Cell Sci. 1994 Mar;107 ( Pt 3):635-44 PMID: 8006077
  7. Sequences controlling transcription of the Chlamydomonas reinhardtii beta 2-tubulin gene after deflagellation and during the cell cycle.
    Mol Cell Biol. 1994 Aug;14(8):5165-74 PMID: 8035797
  8. The ancient regulatory-protein family of WD-repeat proteins.
    Nature. 1994 Sep 22;371(6495):297-300 PMID: 8090199
  9. Components of a "dynein regulatory complex" are located at the junction between the radial spokes and the dynein arms in Chlamydomonas flagella.
    J Cell Biol. 1994 Dec;127(5):1311-25 PMID: 7962092
  10. The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly.
    J Cell Biol. 1995 Apr;129(1):169-78 PMID: 7698982
  11. The 78,000-M(r) intermediate chain of Chlamydomonas outer arm dynein is a microtubule-binding protein.
    J Cell Biol. 1995 Oct;131(2):399-409 PMID: 7593167
  12. Interspecies conservation of outer arm dynein intermediate chain sequences defines two intermediate chain subclasses.
    Mol Biol Cell. 1995 Jun;6(6):685-96 PMID: 7579688
  13. Dynein binds to and crossbridges cytoplasmic microtubules.
    Proc Natl Acad Sci U S A. 1979 Nov;76(11):5759-63 PMID: 160555
  14. Multiple alpha- and beta-tubulin genes in Chlamydomonas and regulation of tubulin mRNA levels after deflagellation.
    Cell. 1981 Apr;24(1):81-8 PMID: 6263492
  15. Enhanced graphic matrix analysis of nucleic acid and protein sequences.
    Proc Natl Acad Sci U S A. 1981 Dec;78(12):7665-9 PMID: 6801656
  16. Substructure of the outer dynein arm.
    J Cell Biol. 1982 Dec;95(3):798-815 PMID: 6218174
  17. Structural comparison of purified dynein proteins with in situ dynein arms.
    J Mol Biol. 1984 Dec 25;180(4):1083-118 PMID: 6241263
  18. A motile Chlamydomonas flagellar mutant that lacks outer dynein arms.
    J Cell Biol. 1985 Apr;100(4):1228-34 PMID: 3156867
  19. The substructure of isolated and in situ outer dynein arms of sea urchin sperm flagella.
    J Cell Biol. 1985 Oct;101(4):1400-12 PMID: 2931439
  20. Isolation of Chlamydomonas flagella and flagellar axonemes.
    Methods Enzymol. 1986;134:280-90 PMID: 3821567
  21. Regulation of the synthesis and assembly of ciliary and flagellar proteins during regeneration.
    Annu Rev Cell Biol. 1986;2:517-46 PMID: 3548775
  22. High-pressure liquid chromatography fractionation of Chlamydomonas dynein extracts and characterization of inner-arm dynein subunits.
    J Mol Biol. 1987 Apr 5;194(3):481-94 PMID: 2957507
  23. 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 PMID: 2958145
  24. Three distinct inner dynein arms in Chlamydomonas flagella: molecular composition and location in the axoneme.
    J Cell Biol. 1990 Feb;110(2):379-89 PMID: 2137128
  25. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  26. Localization of an intermediate chain of outer arm dynein by immunoelectron microscopy.
    J Biol Chem. 1990 Nov 15;265(32):19807-11 PMID: 2147183
  27. Two types of Chlamydomonas flagellar mutants missing different components of inner-arm dynein.
    J Cell Biol. 1991 Feb;112(3):441-7 PMID: 1825085
  28. The proximal portion of Chlamydomonas flagella contains a distinct set of inner dynein arms.
    J Cell Biol. 1991 Feb;112(4):701-9 PMID: 1825211
  29. The Mr 78,000 intermediate chain of Chlamydomonas outer arm dynein interacts with alpha-tubulin in situ.
    J Biol Chem. 1991 May 5;266(13):8401-7 PMID: 1827120
  30. Identification of oda6 as a Chlamydomonas dynein mutant by rescue with the wild-type gene.
    J Cell Biol. 1991 May;113(4):835-42 PMID: 1673970
  31. Predicting coiled coils from protein sequences.
    Science. 1991 May 24;252(5009):1162-4 PMID: 2031185
  32. Microtubule binding and translocation by inner dynein arm subtype I1.
    Cell Motil Cytoskeleton. 1991;18(4):258-68 PMID: 1828714
  33. The inner dynein arm complex: compatible images from freeze-etch and thin section methods of microscopy.
    J Cell Sci. 1991 Oct;100 ( Pt 2):319-28 PMID: 1836791
  34. Structural and functional reconstitution of inner dynein arms in Chlamydomonas flagellar axonemes.
    J Cell Biol. 1992 May;117(3):573-81 PMID: 1533396
  35. The Chlamydomonas Dhc1 gene encodes a dynein heavy chain subunit required for assembly of the I1 inner arm complex.
    Mol Biol Cell. 1997 Apr;8(4):607-20 PMID: 9247642
  36. Overexpression of the dynamitin (p50) subunit of the dynactin complex disrupts dynein-dependent maintenance of membrane organelle distribution.
    J Cell Biol. 1997 Oct 20;139(2):469-84 PMID: 9334349
  37. The involvement of the intermediate chain of cytoplasmic dynein in binding the motor complex to membranous organelles of Xenopus oocytes.
    Mol Biol Cell. 1997 Oct;8(10):2077-88 PMID: 9348543
  38. Identification of the t complex-encoded cytoplasmic dynein light chain tctex1 in inner arm I1 supports the involvement of flagellar dyneins in meiotic drive.
    J Cell Biol. 1998 Mar 9;140(5):1137-47 PMID: 9490726
  39. The Chlamydomonas IDA7 locus encodes a 140-kDa dynein intermediate chain required to assemble the I1 inner arm complex.
    Mol Biol Cell. 1998 Dec;9(12):3351-65 PMID: 9843574
  40. The WD-40 repeat.
    FEBS Lett. 1992 Jul 28;307(2):131-4 PMID: 1644165
  41. Homology of the 74-kD cytoplasmic dynein subunit with a flagellar dynein polypeptide suggests an intracellular targeting function.
    J Cell Biol. 1992 Sep;118(5):1133-43 PMID: 1387402
  42. Arrangement of inner dynein arms in wild-type and mutant flagella of Chlamydomonas.
    J Cell Biol. 1992 Sep;118(5):1145-62 PMID: 1387403
  43. Extragenic suppressors of paralyzed flagellar mutations in Chlamydomonas reinhardtii identify loci that alter the inner dynein arms.
    J Cell Biol. 1992 Sep;118(5):1163-76 PMID: 1387404
  44. The light chain p28 associates with a subset of inner dynein arm heavy chains in Chlamydomonas axonemes.
    Mol Biol Cell. 1995 Jun;6(6):697-711 PMID: 7579689
  45. Affinity chromatography demonstrates a direct binding between cytoplasmic dynein and the dynactin complex.
    J Biol Chem. 1995 Dec 1;270(48):28806-11 PMID: 7499404
  46. Cytoplasmic dynein binds dynactin through a direct interaction between the intermediate chains and p150Glued.
    J Cell Biol. 1995 Dec;131(6 Pt 1):1507-16 PMID: 8522607
  47. The structure of the G protein heterotrimer Gi alpha 1 beta 1 gamma 2.
    Cell. 1995 Dec 15;83(6):1047-58 PMID: 8521505
  48. The 2.0 A crystal structure of a heterotrimeric G protein.
    Nature. 1996 Jan 25;379(6563):311-9 PMID: 8552184
  49. Crystal structure of a G-protein beta gamma dimer at 2.1A resolution.
    Nature. 1996 Jan 25;379(6563):369-74 PMID: 8552196
  50. G protein heterodimers: new structures propel new questions.
    Cell. 1996 Jan 26;84(2):175-8 PMID: 8565060
  51. 1.8 A crystal structure of the C-terminal domain of rabbit serum haemopexin.
    Structure. 1995 Jun 15;3(6):551-9 PMID: 8590016
  52. Axonemal dyneins: assembly, organization, and regulation.
    Curr Opin Cell Biol. 1996 Feb;8(1):10-7 PMID: 8791407
  53. Immunogold localisation of the intermediate chain within the protein complex of cytoplasmic dynein.
    J Struct Biol. 1996 Nov-Dec;117(3):227-35 PMID: 8986653
  54. Regulation of flagellar dynein by phosphorylation of a 138-kD inner arm dynein intermediate chain.
    J Cell Biol. 1997 Jan 13;136(1):167-76 PMID: 9008711
  55. Phosphoregulation of an inner dynein arm complex in Chlamydomonas reinhardtii is altered in phototactic mutant strains.
    J Cell Biol. 1997 Jan 13;136(1):177-91 PMID: 9008712
  56. Studies on the eel sperm flagellum. I. The structure of the inner dynein arm complex.
    J Cell Sci. 1997 Jan;110 ( Pt 1):85-94 PMID: 9010787
  57. The Chlamydomonas reinhardtii ODA3 gene encodes a protein of the outer dynein arm docking complex.
    J Cell Biol. 1997 Jun 2;137(5):1069-80 PMID: 9166407
  58. Microtubule stabilization by assembly-promoting microtubule-associated proteins: a repeat performance.
    Cell Motil Cytoskeleton. 1992;23(4):236-43 PMID: 1477887
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.9.12.3335
Published
1998-12-00
Pages
3335-49
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC25632
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.9.12.3335 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2