Abstract
An antiserum that recognizes the heavy chain of Drosophila kinesin was used to isolate Drosophila cDNA clones. Immunoblot analysis of the proteolytic fragments of the protein produced by one of the cDNA clones has demonstrated that the cDNA clones encode the heavy chain of Drosophila kinesin. The in vitro-synthesized product of the largest cDNA comigrates with Drosophila kinesin heavy chain on NaDodSO4/polyacrylamide gels and binds to taxol-stabilized microtubules in the presence of the nonhydrolyzable analogue of ATP, 5'-adenylyl imidodiphosphate, but not in the presence of ATP or 0.1 M KCl. Analysis of the cDNA clones suggests that there is a single gene encoding kinesin heavy chain in Drosophila located at polytene chromosome position 53A. However, Southern hybridization analyses suggest the presence of related sequences in the Drosophila genome.
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- Bonner J. J., Pardue M. L. Ecdysone-stimulated RNA synthesis in imaginal discs of Drosophila melanogaster. Assay by in situ hybridization. Chromosoma. 1976 Oct 12;58(1):87–99. doi: 10.1007/BF00293443. [DOI] [PubMed] [Google Scholar]
- Brady S. T. A novel brain ATPase with properties expected for the fast axonal transport motor. Nature. 1985 Sep 5;317(6032):73–75. doi: 10.1038/317073a0. [DOI] [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]
- Crowl R., Seamans C., Lomedico P., McAndrew S. Versatile expression vectors for high-level synthesis of cloned gene products in Escherichia coli. Gene. 1985;38(1-3):31–38. doi: 10.1016/0378-1119(85)90200-8. [DOI] [PubMed] [Google Scholar]
- Goldstein L. S., Laymon R. A., McIntosh J. R. A microtubule-associated protein in Drosophila melanogaster: identification, characterization, and isolation of coding sequences. J Cell Biol. 1986 Jun;102(6):2076–2087. doi: 10.1083/jcb.102.6.2076. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Itoh N., Slemmon J. R., Hawke D. H., Williamson R., Morita E., Itakura K., Roberts E., Shively J. E., Crawford G. D., Salvaterra P. M. Cloning of Drosophila choline acetyltransferase cDNA. Proc Natl Acad Sci U S A. 1986 Jun;83(11):4081–4085. doi: 10.1073/pnas.83.11.4081. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackson R. J., Hunt T. Preparation and use of nuclease-treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA. Methods Enzymol. 1983;96:50–74. doi: 10.1016/s0076-6879(83)96008-1. [DOI] [PubMed] [Google Scholar]
- Kuznetsov S. A., Gelfand V. I. Bovine brain kinesin is a microtubule-activated ATPase. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8530–8534. doi: 10.1073/pnas.83.22.8530. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leslie R. J., Hird R. B., Wilson L., McIntosh J. R., Scholey J. M. Kinesin is associated with a nonmicrotubule component of sea urchin mitotic spindles. Proc Natl Acad Sci U S A. 1987 May;84(9):2771–2775. doi: 10.1073/pnas.84.9.2771. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'hare K., Levis R., Rubin G. M. Transcription of the white locus in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6917–6921. doi: 10.1073/pnas.80.22.6917. [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]
- Scholey J. M., Porter M. E., Grissom P. M., McIntosh J. R. Identification of kinesin in sea urchin eggs, and evidence for its localization in the mitotic spindle. Nature. 1985 Dec 5;318(6045):483–486. doi: 10.1038/318483a0. [DOI] [PubMed] [Google Scholar]
- Shepard H. M., Yelverton E., Goeddel D. V. Increased synthesis in E. coli of fibroblast and leukocyte interferons through alterations in ribosome binding sites. DNA. 1982;1(2):125–131. doi: 10.1089/dna.1.1982.1.125. [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., Schnapp B. J., Mitchison T., Steuer E., Reese T. S., Sheetz M. P. Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro. Cell. 1985 Dec;43(3 Pt 2):623–632. doi: 10.1016/0092-8674(85)90234-x. [DOI] [PubMed] [Google Scholar]
- Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]