Abstract
The microtubule motor cytoplasmic dynein has been implicated in a variety of intracellular transport processes. We previously identified and characterized the Drosophila gene Dhc64C, which encodes a cytoplasmic dynein heavy chain. To investigate the function of the cytoplasmic dynein motor, we initiated a mutational analysis of the Dhc64C dynein gene. A small deletion that removes the chromosomal region containing the heavy chain gene was used to isolate EMS-induced lethal mutations that define at least eight essential genes in the region. Germline transformation with a Dhc64C transgene rescued 16 mutant alleles in the single complementation group that identifies the dynein heavy chain gene. All 16 alleles were hemizygous lethal, which demonstrates that the cytoplasmic dynein heavy chain gene Dhc64C is essential for Drosophila development. Furthermore, our failure to recover somatic clones of cells homozygous for a Dhc64C mutation indicates that cytoplasmic dynein function is required for cell viability in several Drosophila tissues. The intragenic complementation of dynein alleles reveals multiple mutant phenotypes including male and/or female sterility, bristle defects, and defects in eye development.
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- Brizuela B. J., Elfring L., Ballard J., Tamkun J. W., Kennison J. A. Genetic analysis of the brahma gene of Drosophila melanogaster and polytene chromosome subdivisions 72AB. Genetics. 1994 Jul;137(3):803–813. doi: 10.1093/genetics/137.3.803. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CRICK F. H., ORGEL L. E. THE THEORY OF INTER-ALLELIC COMPLEMENTATION. J Mol Biol. 1964 Jan;8:161–165. doi: 10.1016/s0022-2836(64)80156-x. [DOI] [PubMed] [Google Scholar]
- Chovnick A. Intragenic recombination in Drosophila: the rosy locus. Genetics. 1989 Dec;123(4):621–624. doi: 10.1093/genetics/123.4.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crosby M. A., Meyerowitz E. M. Lethal mutations flanking the 68C glue gene cluster on chromosome 3 of Drosophila melanogaster. Genetics. 1986 Apr;112(4):785–802. doi: 10.1093/genetics/112.4.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dillman J. F., 3rd, Pfister K. K. Differential phosphorylation in vivo of cytoplasmic dynein associated with anterogradely moving organelles. J Cell Biol. 1994 Dec;127(6 Pt 1):1671–1681. doi: 10.1083/jcb.127.6.1671. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eshel D., Urrestarazu L. A., Vissers S., Jauniaux J. C., van Vliet-Reedijk J. C., Planta R. J., Gibbons I. R. Cytoplasmic dynein is required for normal nuclear segregation in yeast. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11172–11176. doi: 10.1073/pnas.90.23.11172. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gatti M., Baker B. S. Genes controlling essential cell-cycle functions in Drosophila melanogaster. Genes Dev. 1989 Apr;3(4):438–453. doi: 10.1101/gad.3.4.438. [DOI] [PubMed] [Google Scholar]
- Gibbons B. H., Asai D. J., Tang W. J., Hays T. S., Gibbons I. R. Phylogeny and expression of axonemal and cytoplasmic dynein genes in sea urchins. Mol Biol Cell. 1994 Jan;5(1):57–70. doi: 10.1091/mbc.5.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gill S. R., Schroer T. A., Szilak I., Steuer E. R., Sheetz M. P., Cleveland D. W. Dynactin, a conserved, ubiquitously expressed component of an activator of vesicle motility mediated by cytoplasmic dynein. J Cell Biol. 1991 Dec;115(6):1639–1650. doi: 10.1083/jcb.115.6.1639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Golic K. G. Site-specific recombination between homologous chromosomes in Drosophila. Science. 1991 May 17;252(5008):958–961. doi: 10.1126/science.2035025. [DOI] [PubMed] [Google Scholar]
- Hays T. S., Porter M. E., McGrail M., Grissom P., Gosch P., Fuller M. T., McIntosh J. R. A cytoplasmic dynein motor in Drosophila: identification and localization during embryogenesis. J Cell Sci. 1994 Jun;107(Pt 6):1557–1569. doi: 10.1242/jcs.107.6.1557. [DOI] [PubMed] [Google Scholar]
- Holzbaur E. L., Hammarback J. A., Paschal B. M., Kravit N. G., Pfister K. K., Vallee R. B. Homology of a 150K cytoplasmic dynein-associated polypeptide with the Drosophila gene Glued. Nature. 1991 Jun 13;351(6327):579–583. doi: 10.1038/351579a0. [DOI] [PubMed] [Google Scholar]
- Holzbaur E. L., Vallee R. B. DYNEINS: molecular structure and cellular function. Annu Rev Cell Biol. 1994;10:339–372. doi: 10.1146/annurev.cb.10.110194.002011. [DOI] [PubMed] [Google Scholar]
- Karess R. E., Rubin G. M. Analysis of P transposable element functions in Drosophila. Cell. 1984 Aug;38(1):135–146. doi: 10.1016/0092-8674(84)90534-8. [DOI] [PubMed] [Google Scholar]
- King S. M., Patel-King R. S. The M(r) = 8,000 and 11,000 outer arm dynein light chains from Chlamydomonas flagella have cytoplasmic homologues. J Biol Chem. 1995 May 12;270(19):11445–11452. doi: 10.1074/jbc.270.19.11445. [DOI] [PubMed] [Google Scholar]
- Klemenz R., Weber U., Gehring W. J. The white gene as a marker in a new P-element vector for gene transfer in Drosophila. Nucleic Acids Res. 1987 May 26;15(10):3947–3959. doi: 10.1093/nar/15.10.3947. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klämbt C., Jacobs J. R., Goodman C. S. The midline of the Drosophila central nervous system: a model for the genetic analysis of cell fate, cell migration, and growth cone guidance. Cell. 1991 Feb 22;64(4):801–815. doi: 10.1016/0092-8674(91)90509-w. [DOI] [PubMed] [Google Scholar]
- Leptin M., Grunewald B. Cell shape changes during gastrulation in Drosophila. Development. 1990 Sep;110(1):73–84. doi: 10.1242/dev.110.1.73. [DOI] [PubMed] [Google Scholar]
- Li M., McGrail M., Serr M., Hays T. S. Drosophila cytoplasmic dynein, a microtubule motor that is asymmetrically localized in the oocyte. J Cell Biol. 1994 Sep;126(6):1475–1494. doi: 10.1083/jcb.126.6.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li Y. Y., Yeh E., Hays T., Bloom K. Disruption of mitotic spindle orientation in a yeast dynein mutant. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10096–10100. doi: 10.1073/pnas.90.21.10096. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lye R. J., Wilson R. K., Waterston R. H. Genomic structure of a cytoplasmic dynein heavy chain gene from the nematode Caenorhabditis elegans. Cell Motil Cytoskeleton. 1995;32(1):26–36. doi: 10.1002/cm.970320104. [DOI] [PubMed] [Google Scholar]
- McGrail M., Gepner J., Silvanovich A., Ludmann S., Serr M., Hays T. S. Regulation of cytoplasmic dynein function in vivo by the Drosophila Glued complex. J Cell Biol. 1995 Oct;131(2):411–425. doi: 10.1083/jcb.131.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mikami A., Paschal B. M., Mazumdar M., Vallee R. B. Molecular cloning of the retrograde transport motor cytoplasmic dynein (MAP 1C). Neuron. 1993 May;10(5):787–796. doi: 10.1016/0896-6273(93)90195-w. [DOI] [PubMed] [Google Scholar]
- Overton J. The fine structure of developing bristles in wild type and mutant Drosophila melanogaster. J Morphol. 1967 Aug;122(4):367–379. doi: 10.1002/jmor.1051220406. [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]
- Patel N. H., Schafer B., Goodman C. S., Holmgren R. The role of segment polarity genes during Drosophila neurogenesis. Genes Dev. 1989 Jun;3(6):890–904. doi: 10.1101/gad.3.6.890. [DOI] [PubMed] [Google Scholar]
- Pfarr C. M., Coue M., Grissom P. M., Hays T. S., Porter M. E., McIntosh J. R. Cytoplasmic dynein is localized to kinetochores during mitosis. Nature. 1990 May 17;345(6272):263–265. doi: 10.1038/345263a0. [DOI] [PubMed] [Google Scholar]
- Porter M. E., Johnson K. A. Dynein structure and function. Annu Rev Cell Biol. 1989;5:119–151. doi: 10.1146/annurev.cb.05.110189.001003. [DOI] [PubMed] [Google Scholar]
- Rasmusson K., Serr M., Gepner J., Gibbons I., Hays T. S. A family of dynein genes in Drosophila melanogaster. Mol Biol Cell. 1994 Jan;5(1):45–55. doi: 10.1091/mbc.5.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Renfranz P. J., Benzer S. Monoclonal antibody probes discriminate early and late mutant defects in development of the Drosophila retina. Dev Biol. 1989 Dec;136(2):411–429. doi: 10.1016/0012-1606(89)90267-4. [DOI] [PubMed] [Google Scholar]
- Schroer T. A., Sheetz M. P. Two activators of microtubule-based vesicle transport. J Cell Biol. 1991 Dec;115(5):1309–1318. doi: 10.1083/jcb.115.5.1309. [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]
- Schroer T. A. Structure, function and regulation of cytoplasmic dynein. Curr Opin Cell Biol. 1994 Feb;6(1):69–73. doi: 10.1016/0955-0674(94)90118-x. [DOI] [PubMed] [Google Scholar]
- Simmons M. J., Johnson N. A., Fahey T. M., Nellett S. M., Raymond J. D. High mutability in male hybrids of Drosophila melanogaster. Genetics. 1980 Oct;96(2):479–480. doi: 10.1093/genetics/96.2.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steuer E. R., Wordeman L., Schroer T. A., Sheetz M. P. Localization of cytoplasmic dynein to mitotic spindles and kinetochores. Nature. 1990 May 17;345(6272):266–268. doi: 10.1038/345266a0. [DOI] [PubMed] [Google Scholar]
- Tilney L. G., Tilney M. S., Guild G. M. F actin bundles in Drosophila bristles. I. Two filament cross-links are involved in bundling. J Cell Biol. 1995 Aug;130(3):629–638. doi: 10.1083/jcb.130.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vaisberg E. A., Koonce M. P., McIntosh J. R. Cytoplasmic dynein plays a role in mammalian mitotic spindle formation. J Cell Biol. 1993 Nov;123(4):849–858. doi: 10.1083/jcb.123.4.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D., Schnapp B. J., Reese T. S., Sheetz M. P. Organelle, bead, and microtubule translocations promoted by soluble factors from the squid giant axon. Cell. 1985 Mar;40(3):559–569. doi: 10.1016/0092-8674(85)90204-1. [DOI] [PubMed] [Google Scholar]
- Walker R. A., Sheetz M. P. Cytoplasmic microtubule-associated motors. Annu Rev Biochem. 1993;62:429–451. doi: 10.1146/annurev.bi.62.070193.002241. [DOI] [PubMed] [Google Scholar]
- Xiang X., Beckwith S. M., Morris N. R. Cytoplasmic dynein is involved in nuclear migration in Aspergillus nidulans. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2100–2104. doi: 10.1073/pnas.91.6.2100. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Xu T., Rubin G. M. Analysis of genetic mosaics in developing and adult Drosophila tissues. Development. 1993 Apr;117(4):1223–1237. doi: 10.1242/dev.117.4.1223. [DOI] [PubMed] [Google Scholar]