Home LiteratureArticle Details
PMID: 10459015 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas.

The Journal of cell biology ·Vol. 146 ·No. 4 ·1999-08-23 ·Pages 801-18

Myster SH, Knott JA, Wysocki KM, O'Toole E, Porter ME

Abstract

Flagellar motility is generated by the activity of multiple dynein motors, but the specific role of each dynein heavy chain (Dhc) is largely unknown, and the mechanism by which the different Dhcs are targeted to their unique locations is also poorly understood. We report here the complete nucleotide sequence of the Chlamydomonas Dhc1 gene and the corresponding deduced amino acid sequence of the 1alpha Dhc of the I1 inner dynein arm. The 1alpha Dhc is similar to other axonemal Dhcs, but two additional phosphate binding motifs (P-loops) have been identified in the NH(2)- and COOH-terminal regions. Because mutations in Dhc1 result in motility defects and loss of the I1 inner arm, a series of Dhc1 transgenes were used to rescue the mutant phenotypes. Motile cotransformants that express either full-length or truncated 1alpha Dhcs were recovered. The truncated 1alpha Dhc fragments lacked the dynein motor domain, but still assembled with the 1beta Dhc and other I1 subunits into partially functional complexes at the correct axoneme location. Analysis of the transformants has identified the site of the 1alpha motor domain in the I1 structure and further revealed the role of the 1alpha Dhc in flagellar motility and phototactic behavior.

MeSH Terms
Amino Acid Sequence Animals Chlamydomonas/physiology Cosmids/genetics Dyneins/chemistry,genetics,metabolism Flagella/chemistry,physiology Genetic Complementation Test Molecular Motor Proteins/chemistry,genetics,metabolism Molecular Sequence Data Movement Mutation Phenotype Protein Structure, Secondary RNA, Messenger/analysis,genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Transformation, Genetic Transgenes/genetics
Chemicals
Molecular Motor Proteins RNA, Messenger Dyneins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Myster S H
Department of Genetics, Cell Biology, and Development, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
Knott J A
Wysocki K M
O'Toole E
Porter M E
References (71)
71 references, click to expand
  1. Brain cytoplasmic and flagellar outer arm dyneins share a highly conserved Mr 8,000 light chain.
    J Biol Chem. 1996 Aug 9;271(32):19358-66 PMID: 8702622
  2. Beat frequency difference between the two flagella of Chlamydomonas depends on the attachment site of outer dynein arms on the outer-doublet microtubules.
    Cell Motil Cytoskeleton. 1997;36(1):68-75 PMID: 8986378
  3. Purification and characterization of Chlamydomonas flagellar dyneins.
    Methods Enzymol. 1986;134:291-306 PMID: 3029544
  4. Microtubule binding and translocation by inner dynein arm subtype I1.
    Cell Motil Cytoskeleton. 1991;18(4):258-68 PMID: 1828714
  5. Protein-protein interactions in the 18S ATPase of Chlamydomonas outer dynein arms.
    Cell Motil Cytoskeleton. 1986;6(5):510-20 PMID: 2947699
  6. Flagellar photoresponses of ptx1, a nonphototactic mutant of Chlamydomonas.
    Cell Motil Cytoskeleton. 1997;37(2):111-9 PMID: 9186008
  7. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  8. Overexpression of cytoplasmic dynein's globular head causes a collapse of the interphase microtubule network in Dictyostelium.
    Mol Biol Cell. 1996 Jun;7(6):935-48 PMID: 8816999
  9. The DHC1b (DHC2) isoform of cytoplasmic dynein is required for flagellar assembly.
    J Cell Biol. 1999 Feb 8;144(3):473-81 PMID: 9971742
  10. Cytoplasmic dynein heavy chain is an essential gene product in Dictyostelium.
    Cell Motil Cytoskeleton. 1998;39(1):63-72 PMID: 9453714
  11. The genetics and cytology of Chlamydomonas.
    Annu Rev Microbiol. 1960;14:197-216 PMID: 13761500
  12. The role of the dynein stalk in cytoplasmic and flagellar motility.
    Eur Biophys J. 1998;27(5):466-73 PMID: 9760728
  13. The sup-pf-2 mutations of Chlamydomonas alter the activity of the outer dynein arms by modification of the gamma-dynein heavy chain.
    J Cell Biol. 1996 Dec;135(6 Pt 2):1853-65 PMID: 8991096
  14. Submicromolar levels of calcium control the balance of beating between the two flagella in demembranated models of Chlamydomonas.
    J Cell Biol. 1984 Jan;98(1):97-107 PMID: 6707098
  15. Phylogeny and expression of axonemal and cytoplasmic dynein genes in sea urchins.
    Mol Biol Cell. 1994 Jan;5(1):57-70 PMID: 8186465
  16. An extended microtubule-binding structure within the dynein motor domain.
    Nature. 1997 Dec 11;390(6660):636-9 PMID: 9403697
  17. 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
  18. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold.
    EMBO J. 1982;1(8):945-51 PMID: 6329717
  19. The dynein gene family in Chlamydomonas reinhardtii.
    Genetics. 1996 Oct;144(2):569-85 PMID: 8889521
  20. Mutations at twelve independent loci result in absence of outer dynein arms in Chylamydomonas reinhardtii.
    J Cell Biol. 1988 Dec;107(6 Pt 1):2253-8 PMID: 2974040
  21. The CRY1 gene in Chlamydomonas reinhardtii: structure and use as a dominant selectable marker for nuclear transformation.
    Mol Cell Biol. 1994 Jun;14(6):4011-9 PMID: 8196640
  22. Chlamydomonas phototaxis.
    Trends Cell Biol. 1993 Nov;3(11):403-8 PMID: 14731659
  23. Axonemal dyneins: assembly, organization, and regulation.
    Curr Opin Cell Biol. 1996 Feb;8(1):10-7 PMID: 8791407
  24. The dynein genes of Paramecium tetraurelia: the structure and expression of the ciliary beta and cytoplasmic heavy chains.
    Mol Biol Cell. 1995 Nov;6(11):1549-62 PMID: 8589455
  25. Gene isolation through genomic complementation using an indexed library of Chlamydomonas reinhardtii DNA.
    Plant Mol Biol. 1994 Feb;24(4):663-72 PMID: 8155885
  26. The molecular anatomy of dynein.
    Essays Biochem. 2000;35:75-87 PMID: 12471891
  27. Codon usage tabulated from the international DNA sequence databases.
    Nucleic Acids Res. 1997 Jan 1;25(1):244-5 PMID: 9016546
  28. Prediction and analysis of coiled-coil structures.
    Methods Enzymol. 1996;266:513-25 PMID: 8743703
  29. The bop2-1 mutation reveals radial asymmetry in the inner dynein arm region of Chlamydomonas reinhardtii.
    J Cell Biol. 1994 Sep;126(5):1255-66 PMID: 8063862
  30. 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
  31. 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
  32. 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
  33. A motile Chlamydomonas flagellar mutant that lacks outer dynein arms.
    J Cell Biol. 1985 Apr;100(4):1228-34 PMID: 3156867
  34. 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
  35. Two types of Chlamydomonas flagellar mutants missing different components of inner-arm dynein.
    J Cell Biol. 1991 Feb;112(3):441-7 PMID: 1825085
  36. 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
  37. A Chlamydomonas outer arm dynein mutant with a truncated beta heavy chain.
    J Cell Biol. 1993 Aug;122(3):653-61 PMID: 8335691
  38. AAA+: A class of chaperone-like ATPases associated with the assembly, operation, and disassembly of protein complexes.
    Genome Res. 1999 Jan;9(1):27-43 PMID: 9927482
  39. Identification of a microtubule-binding domain in a cytoplasmic dynein heavy chain.
    J Biol Chem. 1997 Aug 8;272(32):19714-8 PMID: 9242627
  40. Cloning of flagellar genes in Chlamydomonas reinhardtii by DNA insertional mutagenesis.
    Genetics. 1993 Oct;135(2):375-84 PMID: 8244002
  41. 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
  42. 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
  43. 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
  44. Complementation of a Chlamydomonas reinhardtii mutant using a genomic cosmid library.
    Plant Mol Biol. 1994 Feb;24(3):533-7 PMID: 8123795
  45. High-frequency nuclear transformation of Chlamydomonas reinhardtii.
    Proc Natl Acad Sci U S A. 1990 Feb;87(3):1228-32 PMID: 2105499
  46. A Chlamydomonas homologue of the putative murine t complex distorter Tctex-2 is an outer arm dynein light chain.
    J Cell Biol. 1997 Jun 2;137(5):1081-90 PMID: 9166408
  47. Cell and molecular biology of flagellar dyneins.
    Int Rev Cytol. 1994;155:141-80 PMID: 7860213
  48. Dynein from Dictyostelium: primary structure comparisons between a cytoplasmic motor enzyme and flagellar dynein.
    J Cell Biol. 1992 Dec;119(6):1597-604 PMID: 1469051
  49. 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
  50. 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
  51. Sequence analysis of the Chlamydomonas alpha and beta dynein heavy chain genes.
    J Cell Sci. 1994 Mar;107 ( Pt 3):635-44 PMID: 8006077
  52. ida4-1, ida4-2, and ida4-3 are intron splicing mutations affecting the locus encoding p28, a light chain of Chlamydomonas axonemal inner dynein arms.
    Mol Biol Cell. 1995 Jun;6(6):713-23 PMID: 7579690
  53. Sequence analysis of the Chlamydomonas reinhardtii flagellar alpha dynein gene.
    Cell Motil Cytoskeleton. 1997;37(2):120-6 PMID: 9186009
  54. 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
  55. Rescue of a paralyzed-flagella mutant of Chlamydomonas by transformation.
    Proc Natl Acad Sci U S A. 1990 Aug;87(15):5739-43 PMID: 2377611
  56. Mutations in the SUP-PF-1 locus of Chlamydomonas reinhardtii identify a regulatory domain in the beta-dynein heavy chain.
    J Cell Biol. 1994 Sep;126(6):1495-507 PMID: 8089181
  57. 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
  58. 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
  59. PF16 encodes a protein with armadillo repeats and localizes to a single microtubule of the central apparatus in Chlamydomonas flagella.
    J Cell Biol. 1996 Feb;132(3):359-70 PMID: 8636214
  60. 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
  61. A Chlamydomonas outer arm dynein mutant missing the alpha heavy chain.
    J Cell Biol. 1991 May;113(3):615-22 PMID: 1673127
  62. Arrangement of inner dynein arms in wild-type and mutant flagella of Chlamydomonas.
    J Cell Biol. 1992 Sep;118(5):1145-62 PMID: 1387403
  63. Cytoplasmic dynein heavy chain 1b is required for flagellar assembly in Chlamydomonas.
    Mol Biol Cell. 1999 Mar;10(3):693-712 PMID: 10069812
  64. The argininosuccinate lyase gene of Chlamydomonas reinhardtii: an important tool for nuclear transformation and for correlating the genetic and molecular maps of the ARG7 locus.
    EMBO J. 1989 Oct;8(10):2803-9 PMID: 2583083
  65. Dynein family of motor proteins: present status and future questions.
    Cell Motil Cytoskeleton. 1995;32(2):136-44 PMID: 8681396
  66. ptx1, a nonphototactic mutant of Chlamydomonas, lacks control of flagellar dominance.
    J Cell Biol. 1993 Feb;120(3):733-41 PMID: 8425899
  67. Isolation of Chlamydomonas flagella and flagellar axonemes.
    Methods Enzymol. 1986;134:280-90 PMID: 3821567
  68. 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
  69. Transformation of chloroplast ribosomal RNA genes in Chlamydomonas: molecular and genetic characterization of integration events.
    Genetics. 1990 Dec;126(4):875-88 PMID: 1981764
  70. 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
  71. Predicting coiled coils from protein sequences.
    Science. 1991 May 24;252(5009):1162-4 PMID: 2031185
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.146.4.801
Published
1999-08-23
Pages
801-18
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2156140
Subset
IM
Grants
NIGMS NIH HHS · GM 55667 · United States
NCRR NIH HHS · RR00592 · United States
Databases
GENBANK
AJ243806
External Links
PubMed source DOI: 10.1083/jcb.146.4.801 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