The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication
The duplication and ninefold symmetry of the Drosophila centriole requires that the cartwheel molecule, Sas6, physically associates with Gorab, a trans-Golgi component. How Gorab achieves these disparate associations is unclear. Here, we use hydrogen–deuterium exchange mass spectrometry to define Go...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2021-03-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/57241 |
| _version_ | 1797991944352169984 |
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| author | Agnieszka Fatalska Emma Stepinac Magdalena Richter Levente Kovacs Zbigniew Pietras Martin Puchinger Gang Dong Michal Dadlez David M Glover |
| author_facet | Agnieszka Fatalska Emma Stepinac Magdalena Richter Levente Kovacs Zbigniew Pietras Martin Puchinger Gang Dong Michal Dadlez David M Glover |
| author_sort | Agnieszka Fatalska |
| collection | DOAJ |
| description | The duplication and ninefold symmetry of the Drosophila centriole requires that the cartwheel molecule, Sas6, physically associates with Gorab, a trans-Golgi component. How Gorab achieves these disparate associations is unclear. Here, we use hydrogen–deuterium exchange mass spectrometry to define Gorab’s interacting surfaces that mediate its subcellular localization. We identify a core stabilization sequence within Gorab’s C-terminal coiled-coil domain that enables homodimerization, binding to Rab6, and thereby trans-Golgi localization. By contrast, part of the Gorab monomer’s coiled-coil domain undergoes an antiparallel interaction with a segment of the parallel coiled-coil dimer of Sas6. This stable heterotrimeric complex can be visualized by electron microscopy. Mutation of a single leucine residue in Sas6’s Gorab-binding domain generates a Sas6 variant with a sixteenfold reduced binding affinity for Gorab that cannot support centriole duplication. Thus, Gorab dimers at the Golgi exist in equilibrium with Sas6-associated monomers at the centriole to balance Gorab’s dual role. |
| first_indexed | 2024-04-11T08:59:08Z |
| format | Article |
| id | doaj.art-1d59bd1d645a4524aa80a89d89ebe3ce |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T08:59:08Z |
| publishDate | 2021-03-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-1d59bd1d645a4524aa80a89d89ebe3ce2022-12-22T04:32:49ZengeLife Sciences Publications LtdeLife2050-084X2021-03-011010.7554/eLife.57241The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplicationAgnieszka Fatalska0https://orcid.org/0000-0002-1720-4742Emma Stepinac1Magdalena Richter2Levente Kovacs3Zbigniew Pietras4Martin Puchinger5Gang Dong6https://orcid.org/0000-0001-9745-8103Michal Dadlez7David M Glover8Department of Genetics, University of Cambridge, Cambridge, United Kingdom; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States; Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, PolandDepartment of Medical Biochemistry, Max Perutz Labs, Medical University of Vienna, Vienna, AustriaDepartment of Genetics, University of Cambridge, Cambridge, United KingdomDepartment of Genetics, University of Cambridge, Cambridge, United Kingdom; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United StatesInstitute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, PolandDepartment of Structural and Computational Biology, Max Perutz Labs, University of Vienna, Vienna, AustriaDepartment of Medical Biochemistry, Max Perutz Labs, Medical University of Vienna, Vienna, AustriaInstitute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, PolandDepartment of Genetics, University of Cambridge, Cambridge, United Kingdom; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United StatesThe duplication and ninefold symmetry of the Drosophila centriole requires that the cartwheel molecule, Sas6, physically associates with Gorab, a trans-Golgi component. How Gorab achieves these disparate associations is unclear. Here, we use hydrogen–deuterium exchange mass spectrometry to define Gorab’s interacting surfaces that mediate its subcellular localization. We identify a core stabilization sequence within Gorab’s C-terminal coiled-coil domain that enables homodimerization, binding to Rab6, and thereby trans-Golgi localization. By contrast, part of the Gorab monomer’s coiled-coil domain undergoes an antiparallel interaction with a segment of the parallel coiled-coil dimer of Sas6. This stable heterotrimeric complex can be visualized by electron microscopy. Mutation of a single leucine residue in Sas6’s Gorab-binding domain generates a Sas6 variant with a sixteenfold reduced binding affinity for Gorab that cannot support centriole duplication. Thus, Gorab dimers at the Golgi exist in equilibrium with Sas6-associated monomers at the centriole to balance Gorab’s dual role.https://elifesciences.org/articles/57241GorabSas6centriole duplicationRab6Golgi |
| spellingShingle | Agnieszka Fatalska Emma Stepinac Magdalena Richter Levente Kovacs Zbigniew Pietras Martin Puchinger Gang Dong Michal Dadlez David M Glover The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication eLife Gorab Sas6 centriole duplication Rab6 Golgi |
| title | The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication |
| title_full | The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication |
| title_fullStr | The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication |
| title_full_unstemmed | The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication |
| title_short | The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication |
| title_sort | dimeric golgi protein gorab binds to sas6 as a monomer to mediate centriole duplication |
| topic | Gorab Sas6 centriole duplication Rab6 Golgi |
| url | https://elifesciences.org/articles/57241 |
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