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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8009671/ https://www.ncbi.nlm.nih.gov/pubmed/33704067 http://dx.doi.org/10.7554/eLife.57241 |
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author | Fatalska, Agnieszka Stepinac, Emma Richter, Magdalena Kovacs, Levente Pietras, Zbigniew Puchinger, Martin Dong, Gang Dadlez, Michal Glover, David M |
author_facet | Fatalska, Agnieszka Stepinac, Emma Richter, Magdalena Kovacs, Levente Pietras, Zbigniew Puchinger, Martin Dong, Gang Dadlez, Michal Glover, David M |
author_sort | Fatalska, Agnieszka |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-8009671 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-80096712021-03-31 The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication Fatalska, Agnieszka Stepinac, Emma Richter, Magdalena Kovacs, Levente Pietras, Zbigniew Puchinger, Martin Dong, Gang Dadlez, Michal Glover, David M eLife Cell Biology 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. eLife Sciences Publications, Ltd 2021-03-11 /pmc/articles/PMC8009671/ /pubmed/33704067 http://dx.doi.org/10.7554/eLife.57241 Text en © 2021, Fatalska et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Cell Biology Fatalska, Agnieszka Stepinac, Emma Richter, Magdalena Kovacs, Levente Pietras, Zbigniew Puchinger, Martin Dong, Gang Dadlez, Michal Glover, David M The dimeric Golgi protein Gorab binds to Sas6 as a monomer to mediate centriole duplication |
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 | Cell Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8009671/ https://www.ncbi.nlm.nih.gov/pubmed/33704067 http://dx.doi.org/10.7554/eLife.57241 |
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