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.
MeSH Terms
Animals
Base Sequence
Cell Line
Centrosome/pathology,physiology,ultrastructure
Chromosome Mapping
DNA Transposable Elements
Drosophila/cytology,embryology,genetics
Immunohistochemistry
Microscopy, Confocal
Mitosis/genetics
Molecular Sequence Data
Mutation
Nervous System/cytology,embryology,ultrastructure
Neurons/cytology,physiology,ultrastructure
Spindle Apparatus/pathology
Tubulin/genetics,isolation & purification,physiology
Chemicals
DNA Transposable Elements
Tubulin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sunkel C E
Centro de Citologia Experimental, Universidade do Porto, Portugal.
Gomes R
Sampaio P
Perdigão J
González C
References (20)
20 references, click to expand
-
Structures of P transposable elements and their sites of insertion and excision in the Drosophila melanogaster genome.
Cell. 1983 Aug;34(1):25-35
PMID: 6309410
-
Gamma-tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation.
Nature. 1992 Mar 5;356(6364):80-3
PMID: 1538786
-
Nuclear antigens follow different pathways into daughter nuclei during mitosis in early Drosophila embryos.
J Cell Sci. 1986 Jun;82:155-72
PMID: 3098744
-
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):3947-59
PMID: 3108854
-
Insertional mutagenesis of the Drosophila genome with single P elements.
Science. 1988 Mar 4;239(4844):1121-8
PMID: 2830671
-
Cloning of a gene encoding an antigen associated with the centrosome in Drosophila.
J Cell Sci. 1988 Apr;89 ( Pt 4):467-80
PMID: 3143740
-
Genetic applications of an inverse polymerase chain reaction.
Genetics. 1988 Nov;120(3):621-3
PMID: 2852134
-
Identification of gamma-tubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans.
Nature. 1989 Apr 20;338(6217):662-4
PMID: 2649796
-
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):1289-301
PMID: 2194669
-
Gamma-tubulin is present in Drosophila melanogaster and Homo sapiens and is associated with the centrosome.
Cell. 1991 May 31;65(5):817-23
PMID: 1904010
-
Drosophila gamma-tubulin is part of a complex containing two previously identified centrosomal MAPs.
J Cell Biol. 1993 May;121(4):823-35
PMID: 8491775
-
gamma-Tubulin participates in the formation of the midbody during cytokinesis in mammalian cells.
J Cell Sci. 1993 May;105 ( Pt 1):145-56
PMID: 8360269
-
gamma-Tubulin is present in acentriolar MTOCs during early mouse development.
J Cell Sci. 1993 May;105 ( Pt 1):157-66
PMID: 8360270
-
The gamma-tubulin gene of the malaria parasite Plasmodium falciparum.
Mol Biochem Parasitol. 1993 Jul;60(1):27-35
PMID: 8366893
-
Centrosome assembly in vitro: role of gamma-tubulin recruitment in Xenopus sperm aster formation.
J Cell Biol. 1994 Jan;124(1-2):19-31
PMID: 8294501
-
Isolation of mip (microtubule-interacting protein) mutations of Aspergillus nidulans.
Mol Cell Biol. 1986 Aug;6(8):2963-8
PMID: 3537728
-
Gamma-tubulin is a highly conserved component of the centrosome.
Cell. 1991 May 31;65(5):825-36
PMID: 1840506
-
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
PMID: 1770000
-
Mutations affecting cell division in Drosophila.
Methods Cell Biol. 1991;35:543-86
PMID: 1664033
-
In vitro reconstitution of centrosome assembly and function: the central role of gamma-tubulin.
Cell. 1994 Feb 25;76(4):623-37
PMID: 8124706