Abstract
We report that in Drosophila, gamma-tubulin is required for the structure as well as the function of microtubule organizing centres (MTOCs). This conclusion is based on the identification and phenotypic characterization of a mutant allele of the gamma-tubulin gene located at region 23C of the polytene chromosome map. This mutation, which we have called gamma-tub23CPl, is caused by the insertion of a P-element within the 5' untranslated leader of the gamma-tubulin transcript. Northern and Western analysis show that gamma-tub23CPl is either a null or a very severe hypomorph as no gamma-tubulin mRNA or protein can be detected in mutant individuals. Visualization of DNA, MTOCs and microtubules by confocal laser scanning microscopy of cells from individuals homozygous for gamma-tub23CPl reveals a series of phenotypic abnormalities. Some of these are similar to those observed after disruption of gamma-tubulin function in other organisms, including mitotic arrest and a dramatic decrease in the number of microtubules, but, in addition, we have observed that mutation in this gene also results in highly abnormal MTOCs which show a variety of shapes and sizes which we never observed in wild type cells. These results show that gamma-tubulin is required for both structural and functional roles in the MTOCs.
Full text
PDF![28](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/8a4b6080e8b1/emboj00025-0038.png)
![29](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/10fa06e8285c/emboj00025-0039.png)
![30](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/4f95894d1f87/emboj00025-0040.png)
![31](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/81265533f3cd/emboj00025-0041.png)
![32](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/340fa215a4bf/emboj00025-0042.png)
![33](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/a6f87fe0fb4e/emboj00025-0043.png)
![34](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/17f2cc9dbb6f/emboj00025-0044.png)
![35](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/03ab6d21b447/emboj00025-0045.png)
![36](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a264/398049/360e1e5d0c6e/emboj00025-0046.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Cooley L., Kelley R., Spradling A. Insertional mutagenesis of the Drosophila genome with single P elements. Science. 1988 Mar 4;239(4844):1121–1128. doi: 10.1126/science.2830671. [DOI] [PubMed] [Google Scholar]
- Frasch M., Glover D. M., Saumweber H. Nuclear antigens follow different pathways into daughter nuclei during mitosis in early Drosophila embryos. J Cell Sci. 1986 Jun;82:155–172. doi: 10.1242/jcs.82.1.155. [DOI] [PubMed] [Google Scholar]
- Félix M. A., Antony C., Wright M., Maro B. Centrosome assembly in vitro: role of gamma-tubulin recruitment in Xenopus sperm aster formation. J Cell Biol. 1994 Jan;124(1-2):19–31. doi: 10.1083/jcb.124.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gatti M., Goldberg M. L. Mutations affecting cell division in Drosophila. Methods Cell Biol. 1991;35:543–586. doi: 10.1016/s0091-679x(08)60587-7. [DOI] [PubMed] [Google Scholar]
- Gueth-Hallonet C., Antony C., Aghion J., Santa-Maria A., Lajoie-Mazenc I., Wright M., Maro B. gamma-Tubulin is present in acentriolar MTOCs during early mouse development. J Cell Sci. 1993 May;105(Pt 1):157–166. doi: 10.1242/jcs.105.1.157. [DOI] [PubMed] [Google Scholar]
- Horio T., Uzawa S., Jung M. K., Oakley B. R., Tanaka K., Yanagida M. The fission yeast gamma-tubulin is essential for mitosis and is localized at microtubule organizing centers. J Cell Sci. 1991 Aug;99(Pt 4):693–700. doi: 10.1242/jcs.99.4.693. [DOI] [PubMed] [Google Scholar]
- Joshi H. C., Palacios M. J., McNamara L., Cleveland D. W. Gamma-tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation. Nature. 1992 Mar 5;356(6364):80–83. doi: 10.1038/356080a0. [DOI] [PubMed] [Google Scholar]
- Julian M., Tollon Y., Lajoie-Mazenc I., Moisand A., Mazarguil H., Puget A., Wright M. gamma-Tubulin participates in the formation of the midbody during cytokinesis in mammalian cells. J Cell Sci. 1993 May;105(Pt 1):145–156. doi: 10.1242/jcs.105.1.145. [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]
- Maessen S., Wesseling J. G., Smits M. A., Konings R. N., Schoenmakers J. G. The gamma-tubulin gene of the malaria parasite Plasmodium falciparum. Mol Biochem Parasitol. 1993 Jul;60(1):27–35. doi: 10.1016/0166-6851(93)90025-s. [DOI] [PubMed] [Google Scholar]
- O'Hare K., Rubin G. M. Structures of P transposable elements and their sites of insertion and excision in the Drosophila melanogaster genome. Cell. 1983 Aug;34(1):25–35. doi: 10.1016/0092-8674(83)90133-2. [DOI] [PubMed] [Google Scholar]
- Oakley B. R., Oakley C. E., Yoon Y., Jung M. K. Gamma-tubulin is a component of the spindle pole body that is essential for microtubule function in Aspergillus nidulans. Cell. 1990 Jun 29;61(7):1289–1301. doi: 10.1016/0092-8674(90)90693-9. [DOI] [PubMed] [Google Scholar]
- Oakley C. E., Oakley B. R. Identification of gamma-tubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans. Nature. 1989 Apr 20;338(6217):662–664. doi: 10.1038/338662a0. [DOI] [PubMed] [Google Scholar]
- Ochman H., Gerber A. S., Hartl D. L. Genetic applications of an inverse polymerase chain reaction. Genetics. 1988 Nov;120(3):621–623. doi: 10.1093/genetics/120.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raff J. W., Kellogg D. R., Alberts B. M. Drosophila gamma-tubulin is part of a complex containing two previously identified centrosomal MAPs. J Cell Biol. 1993 May;121(4):823–835. doi: 10.1083/jcb.121.4.823. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stearns T., Evans L., Kirschner M. Gamma-tubulin is a highly conserved component of the centrosome. Cell. 1991 May 31;65(5):825–836. doi: 10.1016/0092-8674(91)90390-k. [DOI] [PubMed] [Google Scholar]
- Stearns T., Kirschner M. In vitro reconstitution of centrosome assembly and function: the central role of gamma-tubulin. Cell. 1994 Feb 25;76(4):623–637. doi: 10.1016/0092-8674(94)90503-7. [DOI] [PubMed] [Google Scholar]
- Weil C. F., Oakley C. E., Oakley B. R. Isolation of mip (microtubule-interacting protein) mutations of Aspergillus nidulans. Mol Cell Biol. 1986 Aug;6(8):2963–2968. doi: 10.1128/mcb.6.8.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whitfield W. G., Millar S. E., Saumweber H., Frasch M., Glover D. M. Cloning of a gene encoding an antigen associated with the centrosome in Drosophila. J Cell Sci. 1988 Apr;89(Pt 4):467–480. doi: 10.1242/jcs.89.4.467. [DOI] [PubMed] [Google Scholar]
- Zheng Y., Jung M. K., Oakley B. R. Gamma-tubulin is present in Drosophila melanogaster and Homo sapiens and is associated with the centrosome. Cell. 1991 May 31;65(5):817–823. doi: 10.1016/0092-8674(91)90389-g. [DOI] [PubMed] [Google Scholar]