Advertisement
Journal Articles
The dynein genes of Paramecium tetraurelia. Sequences adjacent to the catalytic P-loop identify cytoplasmic and axonemal heavy chain isoforms
D.J. Asai, S.M. Beckwith, K.A. Kandl, H.H. Keating, H. Tjandra, J.D. Forney
Journal of Cell Science 1994 107: 839-847;

Summary

Paramecium tetraurelia is a unicellular organism that utilizes both axonemal and cytoplasmic dyneins. The highly conserved region containing the catalytic P-loop of the dynein heavy chain was amplified by RNA-directed polymerase chain reaction. Eight different P-loop-containing cDNA fragments were cloned. Southern hybridization analysis indicated that each fragment corresponds to a separate dynein gene and that there are at least 12 dynein heavy chain genes expressed in Paramecium. Seven of the eight cloned contain sequence motif A, which is found in axonemal dyneins, and one contains sequence motif B, which is found in the dyneins from cell types that do not have cilia or flagella. Two of the Paramecium dynein genes were further investigated: DHC-6 which contains motif A, and DHC-8 which contains motif B. Additional sequencing of the central portions of these genes showed that DHC-6 most closely matches sea urchin ciliary beta heavy chain and DHC-8 is similar to the cytoplasmic dynein from Dictyostelium. Deciliation of the cells resulted in a substantial increase in the steady state concentration of DHC-6 mRNA but only a small change in DHC-8 mRNA. Antisera were produced against synthetic peptides derived from sequence motifs A and B. Competitive solid-phase binding assays demonstrated that each antiserum was peptide-specific. In western blots, the antiserum to motif A reacted with both ciliary and cytoplasmic dyneins. In contrast, the antiserum to motif B reacted with the cytoplasmic dyneins of Paramecium and bovine brain but did not react with ciliary dynein.(ABSTRACT TRUNCATED AT 250 WORDS)

REFERENCES

    1. Asai D. J.,
    2. Brokaw C. J.,
    3. Thompson W. C. and
    4. Wilson L.
    (1982). Two different monoclonal antibodies to alpha-tubulin inhibit the bending of reactivated sea urchin spermatozoa. Cell Motil 2, 599–614
    OpenUrlCrossRefPubMed
    1. Asai D. J.,
    2. Tang W.-J. Y.,
    3. Ching N. S. and
    4. Gibbons I. R.
    (1991). Cloning and sequencing of the ATP-binding domains of novel isoforms of sea urchin dynein. J. Cell Biol 115, 369–.
    OpenUrl
    1. Beckwith S. M. and
    2. Asai D. J.
    (1993). Ciliary dynein of Paramecium tetraurelia: Photolytic maps of the three heavy chains. Cell Motil. Cytoskel 24, 29–38
    OpenUrlCrossRefPubMed
    1. Beckwith S. M.,
    2. Kandl K. A.,
    3. Forney J. D. and
    4. Asai D. J.
    (1992). Dynein heavy chain isoforms are encoded by separate genes in Paramecium. Mol. Biol. Cell 3, 161–.
    OpenUrl
    1. Brokaw C. J.
    (1991). Microtubule sliding in swimming sperm flagella: Direct and indirect measurements on sea urchin and tunicate spermatozoa. J. Cell Biol 114, 1201–1215
    OpenUrlAbstract/FREE Full Text
    1. Chomczynski P. and
    2. Sacchi N.
    (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem 162, 156–159
    OpenUrlCrossRefPubMedWeb of Science
    1. Cohen J.,
    2. Adoutte A.,
    3. Grandchamp S.,
    4. Houde-Bine L.-M. and
    5. Beisson J.
    (1982). Immunocytochemical study of microtubular structures throughout the cell cycle of Paramecium. Biol. Cell 44, 35–44
    OpenUrl
    1. Corthesy-Theulaz I.,
    2. Pauloin A. and
    3. Pfeffer S. R.
    (1992). Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex. J. Cell Biol 118, 1333–1345
    OpenUrlAbstract/FREE Full Text
    1. Eshel D.,
    2. Urrestarazu L. A.,
    3. Vissers S.,
    4. Jauniaux J.-C.,
    5. van Vliet-Reedijk J. C.,
    6. Planta R. J. and
    7. Gibbons I. R.
    (1993). Cytoplasmic dynein is required for normal nuclear segregation in yeast. Proc. Nat. Acad. Sci. USA 90, 11172–11176
    OpenUrlAbstract/FREE Full Text
    1. Forney J. and
    2. Rodkey K.
    (1992). A repetitive DNA sequence in Paramecium macronuclei is related to thesubunit of G proteins. Nucl. Acids Res 20, 5397–5402
    OpenUrlAbstract/FREE Full Text
    1. Gibbons I. R.
    (1988). Dynein ATPases as microtubule motors. J. Biol. Chem 263, 15837–15840
    OpenUrlFREE Full Text
    1. Gibbons I. R.,
    2. Asai D. J.,
    3. Tang W.-J. Y. and
    4. Gibbons B. H.
    (1992). A cytoplasmic dynein heavy chain in sea urchin embryos. Biol. Cell 76, 303–309
    OpenUrlCrossRefPubMed
    1. Gibbons I. R.,
    2. Gibbons B. H.,
    3. Mocz G. and
    4. Asai D. J.
    (1991). Multiple nucleotide-binding sites in the sequence of dyneinheavy chain. Nature 352, 640–643
    OpenUrlCrossRefPubMed
    1. Gibbons I. R.,
    2. Lee-Eiford A.,
    3. Mocz G.,
    4. Phillipson C. A.,
    5. Tang W.-J. Y. and
    6. Gibbons B. H.
    (1987). Photosensitized cleavage of dynein heavy chains: Cleavage at the ‘V1 site’ by irradiation at 365 nm in the presence of ATP and vanadate. J. Biol. Chem 262, 2780–2786
    OpenUrlAbstract/FREE Full Text
    1. Kagami O. and
    2. Kamiya R.
    (1992). Translocation and rotation of microtubules caused by multiple species of Chlamydomonas inner-arm dynein. J. Cell Sci 103, 653–664
    OpenUrlAbstract
    1. King S. M. and
    2. Witman G. B.
    (1988). Structure of theand heavy chains of the outer arm dynein from Chlamydomonas flagella. Location of epitopes and protease-sensitive sites. J. Biol. Chem 263, 9244–9255
    OpenUrlAbstract/FREE Full Text
    1. Koonce M. P.,
    2. Grissom P. M. and
    3. McIntosh J. R.
    (1992). Dynein from Dictyostelium: Primary structure comparisons between a cytoplasmic motor enzyme and flagellar dynein. J. Cell Biol 119, 1597–1604
    OpenUrlAbstract/FREE Full Text
    1. Larsen J.,
    2. Barakalow K.,
    3. Hamasaki T. and
    4. Satir P.
    (1991). Structural and functional characterization of paramecium dynein: Initial studies. J. Protozool 38, 55–61
    OpenUrlPubMed
    1. Lefebvre P. A. and
    2. Rosenbaum J. L.
    (1986). Regulation of the synthesis and assembly of ciliary and flagellar proteins during regeneration. Annu. Rev. Cell Biol 2, 517–546
    OpenUrlCrossRef
    1. Li Y.-Y.,
    2. Yeh E.,
    3. Hays T. and
    4. Bloom K.
    (1993). Disruption of mitotic spindle orientation in a yeast dynein mutant. Proc. Nat. Acad. Sci. USA 90, 10096–10100
    OpenUrlAbstract/FREE Full Text
    1. Machemer H. and
    2. Ogura A.
    (1979). Ionic conductances of membranes in ciliated and deciliated Paramecium. J. Physiol 296, 49–60
    OpenUrlPubMedWeb of Science
    1. Mikami A.,
    2. Paschal B. M.,
    3. Mazumdar M. and
    4. Vallee R. B.
    (1993). Molecular cloning of the retrograde transport motor cytoplasmic dynein (MAP1C). Neuron 10, 787–796
    OpenUrlCrossRefPubMedWeb of Science
    1. Mitchell D. R. and
    2. Brown K. S.
    (1993). Sequence analysis of two dynein heavy chains from Chlamydomonas. Mol. Biol. Cell 4, 47–.
    OpenUrl
    1. Neely M. D.,
    2. Erickson H. P. and
    3. Boekelheide K.
    (1990). HMW-2, the Sertoli cell cytoplasmic dynein from rat testis, is a dimer composed of nearly identical subunits. J. Biol. Chem 265, 8691–8698
    OpenUrlAbstract/FREE Full Text
    1. Ogawa K.
    (1991). Four ATP-binding sites in the midregion of theheavy chain of dynein. Nature 352, 643–645
    OpenUrlCrossRefPubMedWeb of Science
    1. Pallini V.,
    2. Mencarelli C.,
    3. Bracci L.,
    4. Contorni M.,
    5. Ruggiero P.,
    6. Tiezzi A. and
    7. Manetti R.
    (1983). Cytoplasmic nucleoside-triphosphatases similar to axonemal dynein occur widely in different cell types. J. Submicrosc. Cytol 15, 229–235
    OpenUrlPubMed
    1. Paschal B. M.,
    2. Schpetner H. S. and
    3. Vallee R. B.
    (1987). MAP1C is a microtubule-activated ATPase which translocates microtubules in vitro and has dynein-like properties. J. Cell Biol 105, 1273–1282
    OpenUrlAbstract/FREE Full Text
    1. Pfarr C. M.,
    2. Coue M.,
    3. Grissom P. M.,
    4. Hays T. S.,
    5. Porter M. E. and
    6. McIntosh J. R.
    (1990). Cytoplasmic dynein is localized to kinetochores during mitosis. Nature 345, 263–265
    OpenUrlCrossRefPubMed
    1. Piperno G.
    (1984). Monoclonal antibodies to dynein subunits reveal the existence of cytoplasmic antigens in sea urchin egg. J. Cell Biol 98, 1842–1850
    OpenUrlAbstract/FREE Full Text
    1. Piperno G.,
    2. Ramanis Z.,
    3. Smith E. F. and
    4. Sale W. S.
    (1990). Three distinct inner dynein arms in Chlamydomonas flagella: Molecular composition and location in the axoneme. J. Cell Biol 110, 379–389
    OpenUrlAbstract/FREE Full Text
    1. Porter M.,
    2. Knott J.,
    3. Gardner L.,
    4. Farlow S.,
    5. Myster S. and
    6. Mansanares K.
    (1992). Characterization of the dynein gene family in Chlamydomonas reinhardtii. Mol. Biol. Cell 3, 161–.
    OpenUrl
    1. Satir P.
    (1968). Studies on cilia III. Further studies on the cilium tip and a ‘sliding filament’ model of ciliary motility. J. Cell Biol 39, 77–94
    OpenUrlAbstract/FREE Full Text
    1. Schnapp B. J. and
    2. Reese T. S.
    (1989). Dynein is the motor for retrograde axonal transport of organelles. Proc. Nat. Acad. Sci. USA 86, 1548–1552
    OpenUrlAbstract/FREE Full Text
    1. Schroeder C. C.,
    2. Fok A. K. and
    3. Allen R. D.
    (1990). Vesicle transport alongmicrotubule ribbons and isolation of cytoplasmic dynein from Paramecium. J. Cell Biol 111, 2553–2562
    OpenUrlAbstract/FREE Full Text
    1. Schroer T. A.,
    2. Steuer E. R. and
    3. Sheetz M. P.
    (1989). Cytoplasmic dynein is a minus end-directed motor for membranous organelles. Cell 56, 937–946
    OpenUrlCrossRefPubMedWeb of Science
    1. Shpetner H. S.,
    2. Paschal B. M. and
    3. Vallee R. B.
    (1988). Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP1C). J. Cell Biol 107, 1001–1009
    OpenUrlAbstract/FREE Full Text
    1. Steuer E. R.,
    2. Wordeman L.,
    3. Schroer T. A. and
    4. Sheetz M. P.
    (1990). Localization of cytoplasmic dynein to mitotic spindles and kinetochores. Nature 345, 266–268
    OpenUrlCrossRefPubMed
    1. Travis S. M. and
    2. Nelson D. L.
    (1988). Purification and properties of dyneins from Paramecium cilia. Biochim. Biophys. Acta 966, 73–83
    OpenUrlPubMed
    1. Vaisberg E. A.,
    2. Koonce M. P. and
    3. McIntosh J. R.
    (1993). Cytoplasmic dynein plays a role in mammalian mitotic spindle formation. J. Cell Biol 123, 849–858
    OpenUrlAbstract/FREE Full Text
    1. Vallee R. B.
    (1993). Molecular analysis of the microtubule motor dynein. Proc. Nat. Acad. Sci. USA 90, 8769–8772
    OpenUrlAbstract/FREE Full Text
    1. Verde F.,
    2. Berrez J.-M.,
    3. Antony C. and
    4. Karsenti E.
    (1991). Taxol-induced microtubule asters in mitotic extracts of Xenopus eggs: Requirement for phosphorylated factors and cytoplasmic dynein. J. Cell Biol 112, 1177–1187
    OpenUrlAbstract/FREE Full Text
    1. Wilkerson C. G.,
    2. King S. M. and
    3. Witman G. B.
    (1992). Sequence analysis of multiple dynein heavy chain genes from Chlamydomonas reinhardtii. Mol. Biol. Cell 3, 161–.
    OpenUrl
    1. Wilkerson C. G.,
    2. King S. M. and
    3. Witman G. B.
    (1993). Sequence analysis of thedynein heavy chain from the Chlamydomonas outer arm. Mol. Biol. Cell 4, 47–.
    OpenUrl
    1. Witman G. B.
    (1992). Axonemal dyneins. Curr. Opin. Cell Biol 4, 74–79
    OpenUrlCrossRefPubMed
    1. Wordeman L.,
    2. Steuer E. R.,
    3. Sheetz M. P. and
    4. Mitchison T.
    (1991). Chemical subdomains within the kinetochore domain of isolated CHO mitotic chromosomes. J. Cell Biol 114, 285–294
    OpenUrlAbstract/FREE Full Text
    1. Zhang Z.,
    2. Tanaka Y.,
    3. Nonaka S.,
    4. Aizawa H.,
    5. Kawasaki H.,
    6. Nakata T. and
    7. Hirokawa N.
    (1993). The primary structure of rat brain (cytoplasmic) dynein heavy chain, a cytoplasmic motor enzyme. Proc. Nat. Acad. Sci. USA 90, 7928–7932
    OpenUrlAbstract/FREE Full Text
Back to top

 Download PDF

Email
Journal Articles
The dynein genes of Paramecium tetraurelia. Sequences adjacent to the catalytic P-loop identify cytoplasmic and axonemal heavy chain isoforms
D.J. Asai, S.M. Beckwith, K.A. Kandl, H.H. Keating, H. Tjandra, J.D. Forney
Journal of Cell Science 1994 107: 839-847;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Advertisement

First Person interviews

Photo collage of Chen Yu, Sushmita Chatterjee, Ruiqi Wang and Lotte Vanheer.

Have you seen our First Person interviews with the early-career first authors of our papers? The authors talk about their work in and out of the lab, the journeys that led them to where they are now and the scientists who inspired them along the way. Recently, we caught up with first authors Maitreyi Rathod, Sushmita Chatterjee, Lotte Vanheer, Rachel Furlong, Pamela Adami, Ruiqi Wang and Chen Yu.

Travelling Fellowship – New imaging approach unveils a bigger picture

Highlights from Travelling Fellowship trips

Find out how Pamela Imperadore’s Travelling Fellowship grant from The Company of Biologists took her to Germany, where she used new imaging techniques to investigate the cellular machinery underlying octopus arm regeneration. Don’t miss the next application deadline for 2020 travel, coming up on 29 November. Where will your research take you?

preLights – Meet the preLighters: an interview with Maiko Kitaoka

Maiko Kitaoka

Maiko Kitaoka is a graduate student in the lab of Rebecca Heald at the University of California, Berkeley. Here she studies the cause of chromosome mis-segregation defects in Xenopus hybrids. We caught up with Maiko to discuss her research, science communication, ballet, preprints and more.

Journal Meeting – Cell Dynamics: Host-Pathogen Interface

Registration is now open for the third instalment of the highly successful Cellular Dynamics Meeting Series, and will focus on ‘Host-Pathogen Interface’. The meeting will take place 17-20 May 2020, and further information is available here.

Cell Science at a Glance – Adaptor protein complexes and disease at a glance

A new poster from the Robinson lab summarises what is known about the five adaptor protein complexes and discuss how this helps to explain the clinical features of different genetic disorders.

JCS joins the Review Commons initiative

Journal of Cell Science is pleased to be a part of the new and exciting Review Commons initiative, launched by EMBO and ASAPbio. Streamlining the publishing process, Review Commons enables high-quality peer review to take place before journal submission. Papers submitted to Review Commons will be assessed independently of any journal, focusing solely on the paper’s scientific rigor and merit.