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Gorab is a Golgi protein required for structure and duplication of Drosophila centrioles

Kovacs, Levente and Chao-Chu, Jennifer and Schneider, Sandra and Gottardo, Marco and Tzolovsky, George and Dzhindzhev, Nikola S. and Riparbelli, Maria Giovanna and Callaini, Giuliano and Glover, David M. (2018) Gorab is a Golgi protein required for structure and duplication of Drosophila centrioles. Nature Genetics, 50 (7). pp. 1021-1031. ISSN 1061-4036. PMCID PMC6097609. https://resolver.caltech.edu/CaltechAUTHORS:20200807-114540050

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[img] Image (JPEG) (Supplementary Figure 1: Localization of Gorab in mitotic cells) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 2: Gorab interaction with Sas6) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 3: Sas6 interaction with Gorab) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 4: Localization of V266P mutant Gorab to basal bodies) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 5: C-terminally GFP-tagged wild-type Gorab rescues gorab¹ mutant phenotypes) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 6: Centrosomal localization of human GORAB) - Supplemental Material
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Abstract

We demonstrate that a Drosophila Golgi protein, Gorab, is present not only in the trans-Golgi but also in the centriole cartwheel where, complexed to Sas6, it is required for centriole duplication. In addition to centriole defects, flies lacking Gorab are uncoordinated due to defects in sensory cilia, which lose their nine-fold symmetry. We demonstrate the separation of centriole and Golgi functions of Drosophila Gorab in two ways: first, we have created Gorab variants that are unable to localize to trans-Golgi but can still rescue the centriole and cilia defects of gorab null flies; second, we show that expression of C-terminally tagged Gorab disrupts Golgi functions in cytokinesis of male meiosis, a dominant phenotype overcome by mutations preventing Golgi targeting. Our findings suggest that during animal evolution, a Golgi protein has arisen with a second, apparently independent, role in centriole duplication.


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Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/s41588-018-0149-1DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6097609/PubMed CentralArticle
https://rdcu.be/b58xnPublisherFree ReadCube access
ORCID:
AuthorORCID
Kovacs, Levente0000-0002-3226-3740
Chao-Chu, Jennifer0000-0001-6740-8797
Gottardo, Marco0000-0001-8649-7799
Dzhindzhev, Nikola S.0000-0001-9866-3600
Callaini, Giuliano0000-0003-2252-0309
Glover, David M.0000-0003-0956-0103
Additional Information:© 2018 Nature America. Received 08 May 2017. Accepted 17 April 2018. Published 11 June 2018. Issue Date July 2018. We acknowledge P. Deák (Department of Genetics, University of Szeged, Szeged, Hungary) for his encouragement and support of L.K. and M. Pál (Department of Genetics, University of Szeged, Szeged, Hungary) for injection of CRISPR–Cas9 guide-RNAs for Gorab mutagenesis; and thank S. Chang, K. Oras, and A. Madich (Deparment of Genetics, University of Cambridge, Cambridge, UK) for injection of Gorab transgenes. We also greatly appreciate the advice of M. Richter and A. Fatalska in studies of protein–protein interactions. We thank T. Megraw (Department of Biomedical Sciences, Florida State University, Tallahassee, FL, USA) for GFP-rootletin flies, C. Gonzalez (Institute for Research in Biomedicine, The Barcelona Institute of Science and Technology, Barcelona, Spain) for YFP-centrobin flies, S. Munro (MRC Laboratory of Molecular Biology, Cambridge, UK) for anti-golgin antibodies, K. Raj (Radiation Effects Department, Public Health England, Didcot, UK) for the U2OS^(p53DD) cell line, and J. Debski for advice in mass-spectrometry. D.M.G. is grateful for a Wellcome Investigator Award, which supported this work. The study was initiated with support from Cancer Research UK. Author Contributions: L.K. contributed to planning experiments, fluorescence microscopy, Drosophila genetics, super-resolution microscopy, cell culture work, data analysis, and writing the manuscript; J.C.-C. contributed to antibody generation, mass spectrometry, Drosophila genetics, and fluorescence microscopy; S.S. contributed to mass spectrometry, in vitro interaction studies, protein structure analysis, and cell-culture work; M.G. contributed to electron microscopy; G.T. contributed to super-resolution microscopy; N.S.D. contributed to planning experiments and data analysis; M.G.R. contributed to electron microscopy; G.C. contributed to electron microscopy; and D.M.G. contributed to the conception and supervision of the study, planning experiments, and writing the manuscript. The authors declare no competing interests.
Funders:
Funding AgencyGrant Number
Wellcome TrustUNSPECIFIED
Cancer Research UKUNSPECIFIED
Issue or Number:7
PubMed Central ID:PMC6097609
Record Number:CaltechAUTHORS:20200807-114540050
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200807-114540050
Official Citation:Kovacs, L., Chao-Chu, J., Schneider, S. et al. Gorab is a Golgi protein required for structure and duplication of Drosophila centrioles. Nat Genet 50, 1021–1031 (2018). https://doi.org/10.1038/s41588-018-0149-1
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:104820
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:10 Aug 2020 14:15
Last Modified:10 Aug 2020 14:15

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