Abstract
Membrane-bound organelles move bidirectionally along microtubules in the freshwater ameba, Reticulomyxa. We have examined the nucleotide requirements for transport in a lysed cell model and compared them with kinesin and dynein-driven motility in other systems. Both anterograde and retrograde transport in Reticulomyxa show features characteristic of dynein but not of kinesin-powered movements: organelle transport is reactivated only by ATP and no other nucleoside triphosphates; the Km and Vmax of the ATP-driven movements are similar to values obtained for dynein rather than kinesin-driven movement; and of 15 ATP analogues tested for their ability to promote organelle transport, only 4 of them did. This narrow specificity resembles that of dynein-mediated in vitro transport and is dissimilar to the broad specificity of the kinesin motor (Shimizu, T., K. Furusawa, S. Ohashi, Y. Y. Toyoshima, M. Okuno, F. Malik, and R. D. Vale. 1991. J. Cell Biol. 112: 1189-1197). Remarkably, anterograde and retrograde organelle transport cannot be distinguished at all with respect to nucleotide specificity, kinetics of movement, and the ability to use the ATP analogues. Since the "kinetic fingerprints" of the motors driving transport in opposite directions are indistinguishable, the same type of motor(s) may be involved in the two directions of movement.
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- Brady S. T., Pfister K. K., Bloom G. S. A monoclonal antibody against kinesin inhibits both anterograde and retrograde fast axonal transport in squid axoplasm. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1061–1065. doi: 10.1073/pnas.87.3.1061. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohn S. A., Ingold A. L., Scholey J. M. Correlation between the ATPase and microtubule translocating activities of sea urchin egg kinesin. Nature. 1987 Jul 9;328(6126):160–163. doi: 10.1038/328160a0. [DOI] [PubMed] [Google Scholar]
- Cohn S. A., Ingold A. L., Scholey J. M. Quantitative analysis of sea urchin egg kinesin-driven microtubule motility. J Biol Chem. 1989 Mar 15;264(8):4290–4297. [PubMed] [Google Scholar]
- Dabora S. L., Sheetz M. P. The microtubule-dependent formation of a tubulovesicular network with characteristics of the ER from cultured cell extracts. Cell. 1988 Jul 1;54(1):27–35. doi: 10.1016/0092-8674(88)90176-6. [DOI] [PubMed] [Google Scholar]
- Euteneuer U., Haimo L. T., Schliwa M. Microtubule bundles of Reticulomyxa networks are of uniform polarity. Eur J Cell Biol. 1989 Aug;49(2):373–376. [PubMed] [Google Scholar]
- Euteneuer U., Koonce M. P., Pfister K. K., Schliwa M. An ATPase with properties expected for the organelle motor of the giant amoeba, Reticulomyxa. Nature. 1988 Mar 10;332(6160):176–178. doi: 10.1038/332176a0. [DOI] [PubMed] [Google Scholar]
- Gibbons I. R. Dynein ATPases as microtubule motors. J Biol Chem. 1988 Nov 5;263(31):15837–15840. [PubMed] [Google Scholar]
- Hollenbeck P. J. The distribution, abundance and subcellular localization of kinesin. J Cell Biol. 1989 Jun;108(6):2335–2342. doi: 10.1083/jcb.108.6.2335. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howard J., Hudspeth A. J., Vale R. D. Movement of microtubules by single kinesin molecules. Nature. 1989 Nov 9;342(6246):154–158. doi: 10.1038/342154a0. [DOI] [PubMed] [Google Scholar]
- Huitorel P. From cilia and flagella to intracellular motility and back again: a review of a few aspects of microtubule-based motility. Biol Cell. 1988;63(2):249–258. doi: 10.1016/0248-4900(88)90062-7. [DOI] [PubMed] [Google Scholar]
- Huxley H. E. The mechanism of muscular contraction. Science. 1969 Jun 20;164(3886):1356–1365. doi: 10.1126/science.164.3886.1356. [DOI] [PubMed] [Google Scholar]
- Koonce M. P., Euteneuer U., McDonald K. L., Menzel D., Schliwa M. Cytoskeletal architecture and motility in a giant freshwater amoeba, Reticulomyxa. Cell Motil Cytoskeleton. 1986;6(5):521–533. doi: 10.1002/cm.970060511. [DOI] [PubMed] [Google Scholar]
- Koonce M. P., Schliwa M. Reactivation of organelle movements along the cytoskeletal framework of a giant freshwater ameba. J Cell Biol. 1986 Aug;103(2):605–612. doi: 10.1083/jcb.103.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koonce M. P., Tong J., Euteneuer U., Schliwa M. Active sliding between cytoplasmic microtubules. Nature. 1987 Aug 20;328(6132):737–739. doi: 10.1038/328737a0. [DOI] [PubMed] [Google Scholar]
- Lee-Eiford A., Ow R. A., Gibbons I. R. Specific cleavage of dynein heavy chains by ultraviolet irradiation in the presence of ATP and vanadate. J Biol Chem. 1986 Feb 15;261(5):2337–2342. [PubMed] [Google Scholar]
- Leopold P. L., Snyder R., Bloom G. S., Brady S. T. Nucleotide specificity for the bidirectional transport of membrane-bounded organelles in isolated axoplasm. Cell Motil Cytoskeleton. 1990;15(4):210–219. doi: 10.1002/cm.970150404. [DOI] [PubMed] [Google Scholar]
- Lye R. J., Porter M. E., Scholey J. M., McIntosh J. R. Identification of a microtubule-based cytoplasmic motor in the nematode C. elegans. Cell. 1987 Oct 23;51(2):309–318. doi: 10.1016/0092-8674(87)90157-7. [DOI] [PubMed] [Google Scholar]
- McIntosh J. R., Porter M. E. Enzymes for microtubule-dependent motility. J Biol Chem. 1989 Apr 15;264(11):6001–6004. [PubMed] [Google Scholar]
- Oiwa K., Takahashi K. The force-velocity relationship for microtubule sliding in demembranated sperm flagella of the sea urchin. Cell Struct Funct. 1988 Jun;13(3):193–205. doi: 10.1247/csf.13.193. [DOI] [PubMed] [Google Scholar]
- Paschal B. M., Vallee R. B. Retrograde transport by the microtubule-associated protein MAP 1C. Nature. 1987 Nov 12;330(6144):181–183. doi: 10.1038/330181a0. [DOI] [PubMed] [Google Scholar]
- Pfister K. K., Wagner M. C., Stenoien D. L., Brady S. T., Bloom G. S. Monoclonal antibodies to kinesin heavy and light chains stain vesicle-like structures, but not microtubules, in cultured cells. J Cell Biol. 1989 Apr;108(4):1453–1463. doi: 10.1083/jcb.108.4.1453. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Porter M. E., Scholey J. M., Stemple D. L., Vigers G. P., Vale R. D., Sheetz M. P., McIntosh J. R. Characterization of the microtubule movement produced by sea urchin egg kinesin. J Biol Chem. 1987 Feb 25;262(6):2794–2802. [PubMed] [Google Scholar]
- Pratt M. M. Stable complexes of axoplasmic vesicles and microtubules: protein composition and ATPase activity. J Cell Biol. 1986 Sep;103(3):957–968. doi: 10.1083/jcb.103.3.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rozdzial M. M., Haimo L. T. Reactivated melanophore motility: differential regulation and nucleotide requirements of bidirectional pigment granule transport. J Cell Biol. 1986 Dec;103(6 Pt 2):2755–2764. doi: 10.1083/jcb.103.6.2755. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saxton W. M., Porter M. E., Cohn S. A., Scholey J. M., Raff E. C., McIntosh J. R. Drosophila kinesin: characterization of microtubule motility and ATPase. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1109–1113. doi: 10.1073/pnas.85.4.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schliwa M., van Blerkom J. Structural interaction of cytoskeletal components. J Cell Biol. 1981 Jul;90(1):222–235. doi: 10.1083/jcb.90.1.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schnapp B. J., Reese T. S. Dynein is the motor for retrograde axonal transport of organelles. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1548–1552. doi: 10.1073/pnas.86.5.1548. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schroer T. A., Steuer E. R., Sheetz M. P. Cytoplasmic dynein is a minus end-directed motor for membranous organelles. Cell. 1989 Mar 24;56(6):937–946. doi: 10.1016/0092-8674(89)90627-2. [DOI] [PubMed] [Google Scholar]
- Shimizu T., Furusawa K., Ohashi S., Toyoshima Y. Y., Okuno M., Malik F., Vale R. D. Nucleotide specificity of the enzymatic and motile activities of dynein, kinesin, and heavy meromyosin. J Cell Biol. 1991 Mar;112(6):1189–1197. doi: 10.1083/jcb.112.6.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D., Hotani H. Formation of membrane networks in vitro by kinesin-driven microtubule movement. J Cell Biol. 1988 Dec;107(6 Pt 1):2233–2241. doi: 10.1083/jcb.107.6.2233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D. Intracellular transport using microtubule-based motors. Annu Rev Cell Biol. 1987;3:347–378. doi: 10.1146/annurev.cb.03.110187.002023. [DOI] [PubMed] [Google Scholar]
- Vale R. D., Reese T. S., Sheetz M. P. Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell. 1985 Aug;42(1):39–50. doi: 10.1016/s0092-8674(85)80099-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D., Toyoshima Y. Y. Rotation and translocation of microtubules in vitro induced by dyneins from Tetrahymena cilia. Cell. 1988 Feb 12;52(3):459–469. doi: 10.1016/s0092-8674(88)80038-2. [DOI] [PubMed] [Google Scholar]
- Yano Y., Miki-Noumura T. Sliding velocity between outer doublet microtubules of sea-urchin sperm axonemes. J Cell Sci. 1980 Aug;44:169–186. doi: 10.1242/jcs.44.1.169. [DOI] [PubMed] [Google Scholar]