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Mutation of a Drosophila gamma tubulin ring complex subunit encoded by discs degenerate-4 differentially disrupts centrosomal protein localization

Barbosa, Vitor and Yamamoto, Rochele R. and Henderson, Daryl S. and Glover, David M. (2000) Mutation of a Drosophila gamma tubulin ring complex subunit encoded by discs degenerate-4 differentially disrupts centrosomal protein localization. Genes and Development, 14 (24). pp. 3126-3139. ISSN 0890-9369. PMCID PMC317135. doi:10.1101/gad.182800.

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We have cloned the Drosophila gene discs degenerate-4 (dd4) and find that it encodes a component of the γ-tubulin ring complex (γTuRC) homologous to Spc98 of budding yeast. This provides the first opportunity to study decreased function of a member of the γ-tubulin ring complex, other than γ-tubulin itself, in a metazoan cell. γ-tubulin is no longer at the centrosomes but is dispersed throughout dd4 cells and yet bipolar metaphase spindles do form, although these have a dramatically decreased density of microtubules. Centrosomin (CNN) remains in broad discrete bodies but only at the focused poles of such spindles, whereas Asp (abnormal spindleprotein) is always present at the presumptive minus ends of microtubules, whether or not they are focused. This is consistent with the proposed role of Asp in coordinating the nucleation of mitotic microtubule organizing centers. The centrosome associated protein CP190 is partially lost from the spindle poles in dd4cells supporting a weak interaction with γ-tubulin, and the displaced protein accumulates in the vicinity of chromosomes. Electron microscopy indicates not only that the poles of dd4 cells have irregular amounts of pericentriolar material, but also that they can have abnormal centrioles. In six dd4 cells subjected to serial sectioning centrioles were missing from one of the two poles. This suggests that in addition to its role in nucleating cytoplasmic and spindle microtubules, the γTuRC is also essential to the structure of centrioles and the separation of centrosomes.

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Glover, David M.0000-0003-0956-0103
Additional Information:© 2000 by Cold Spring Harbor Laboratory Press. The Authors acknowledge that six months after the full-issue publication date, the Article will be distributed under a Creative Commons CC-BY-NC License (Attribution-NonCommercial 4.0 International License, Received June 16, 2000; revised version accepted October 24, 2000. We thank the Cancer Research Campaign for supporting this work. V.B. received studentships from the FCT through the Gulbenkian Foundation of Portugal and the UK Medical Research Council. R.Y. received a studentship from CNPq, Brasil. We thank Carmo Avides for help in some of the immunostaining experiments, Yixian Zheng for providing antibody to Dgrip91, and Endre Mathe, Emma Warbrick, and Yutaka Yamamoto for their advice. We thank Abe Schalet and Adelaide Carpenter for providing several mutant strains each. We further thank Adelaide Carpenter for sharing unpublished electron micrographs with us, and for her helpful comments on the manuscript. The publication costs of this article were defrayed in part by payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 USC section 1734 solely to indicate this fact.
Funding AgencyGrant Number
Cancer Research CampaignUNSPECIFIED
Fundação para a Ciência e a Tecnologia (FCT)UNSPECIFIED
Calouste Gulbenkian FoundationUNSPECIFIED
Medical Research Council (UK)UNSPECIFIED
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)UNSPECIFIED
Subject Keywords:γ-TuRC; centrosomes; centrioles; mitosis; spindle; microtubules
Issue or Number:24
PubMed Central ID:PMC317135
Record Number:CaltechAUTHORS:20200807-170353101
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Official Citation:Mutation of a Drosophila gamma tubulin ring complex subunit encoded by discs degenerate-4 differentially disrupts centrosomal protein localization Vitor Barbosa, Rochele R. Yamamoto, Daryl S. Henderson, and David M. Glover Genes Dev. December 15, 2000 14: 3126-3139; doi:10.1101/gad.182800
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:104860
Deposited By: George Porter
Deposited On:19 Aug 2020 22:48
Last Modified:16 Nov 2021 18:37

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