Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published July 2018 | Accepted Version + Supplemental Material
Journal Article Open

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

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.

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.

Attached Files

Accepted Version - emss-77175.pdf

Supplemental Material - 41588_2018_149_Fig10_ESM.jpg

Supplemental Material - 41588_2018_149_Fig11_ESM.jpg

Supplemental Material - 41588_2018_149_Fig12_ESM.jpg

Supplemental Material - 41588_2018_149_Fig13_ESM.jpg

Supplemental Material - 41588_2018_149_Fig8_ESM.jpg

Supplemental Material - 41588_2018_149_Fig9_ESM.jpg

Supplemental Material - 41588_2018_149_MOESM1_ESM.pdf

Supplemental Material - 41588_2018_149_MOESM2_ESM.pdf

Supplemental Material - 41588_2018_149_MOESM3_ESM.pdf

Supplemental Material - 41588_2018_149_MOESM4_ESM.mov

Files

41588_2018_149_MOESM3_ESM.pdf
Files (29.6 MB)
Name Size Download all
md5:9981e1132c08f22165db292bf8515e92
245.8 kB Preview Download
md5:598d8404c94f6045e321ff95d5467b39
77.7 kB Preview Download
md5:f3ae4122ff37b6ec5078c61d0eea5291
23.8 MB Download
md5:cc995c0eb7cb946ef12533619d3b6354
3.8 MB Preview Download
md5:3a3615820e11ac291620d7a1b090a5c0
75.7 kB Preview Download
md5:906775f60db97292dbd98f330d587e41
99.9 kB Preview Download
md5:40047408f044c1a358a99f4f44d822f0
93.2 kB Preview Download
md5:0c9a6faf573e21a022709a450c64fb0c
108.5 kB Preview Download
md5:d692772ddf65af92d78994fd1c4c8bbf
359.9 kB Preview Download
md5:dfb626c54870a7438ab7164630b51951
919.0 kB Preview Download
md5:9fcb7bd2d33ed3d94e2cf8abdf2e7e73
77.9 kB Preview Download

Additional details

Created:
August 19, 2023
Modified:
October 20, 2023