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Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab
The ability to relocalize proteins to defined subcellular locations presents a powerful tool to examine protein-protein interactions that can overcome a tendency of non-targeted exogenous proteins to form inaccessible aggregates. Here, we show that a 24-amino-acid sequence from the Drosophila proapo...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists Ltd
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9836085/ https://www.ncbi.nlm.nih.gov/pubmed/36331102 http://dx.doi.org/10.1242/bio.059545 |
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author | Kovacs, Levente Fatalska, Agnieszka Glover, David M. |
author_facet | Kovacs, Levente Fatalska, Agnieszka Glover, David M. |
author_sort | Kovacs, Levente |
collection | PubMed |
description | The ability to relocalize proteins to defined subcellular locations presents a powerful tool to examine protein-protein interactions that can overcome a tendency of non-targeted exogenous proteins to form inaccessible aggregates. Here, we show that a 24-amino-acid sequence from the Drosophila proapoptotic protein Hid's tail anchor (HTA) domain can target exogenous proteins to the mitochondria in Drosophila cells. We use this HTA tag to target the Drosophila centriole cartwheel protein Sas6 to the mitochondria, and show that both exogenous and endogenous Gorab can be co-recruited from the Golgi to the new mitochondrial site. This accords with our previous observation that monomeric Drosophila Gorab binds Sas6 to become centriole associated with a 50-fold greater affinity than dimeric Gorab binds Rab6 to become localized at the Golgi. Strikingly, Drosophila Sas6 can bind both Drosophila Gorab and its human GORAB ortholog, whereas human SAS6 is unable to bind either GORAB or Gorab. We discuss these findings in relation to the evolutionary conservation of Gorab and the divergence of Sas6, possibly reflecting known differences in persistence of the cartwheel in the centriole duplication cycle of fly and human cells. |
format | Online Article Text |
id | pubmed-9836085 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Company of Biologists Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-98360852023-01-13 Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab Kovacs, Levente Fatalska, Agnieszka Glover, David M. Biol Open Research Article The ability to relocalize proteins to defined subcellular locations presents a powerful tool to examine protein-protein interactions that can overcome a tendency of non-targeted exogenous proteins to form inaccessible aggregates. Here, we show that a 24-amino-acid sequence from the Drosophila proapoptotic protein Hid's tail anchor (HTA) domain can target exogenous proteins to the mitochondria in Drosophila cells. We use this HTA tag to target the Drosophila centriole cartwheel protein Sas6 to the mitochondria, and show that both exogenous and endogenous Gorab can be co-recruited from the Golgi to the new mitochondrial site. This accords with our previous observation that monomeric Drosophila Gorab binds Sas6 to become centriole associated with a 50-fold greater affinity than dimeric Gorab binds Rab6 to become localized at the Golgi. Strikingly, Drosophila Sas6 can bind both Drosophila Gorab and its human GORAB ortholog, whereas human SAS6 is unable to bind either GORAB or Gorab. We discuss these findings in relation to the evolutionary conservation of Gorab and the divergence of Sas6, possibly reflecting known differences in persistence of the cartwheel in the centriole duplication cycle of fly and human cells. The Company of Biologists Ltd 2022-11-18 /pmc/articles/PMC9836085/ /pubmed/36331102 http://dx.doi.org/10.1242/bio.059545 Text en © 2022. Published by The Company of Biologists Ltd https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. |
spellingShingle | Research Article Kovacs, Levente Fatalska, Agnieszka Glover, David M. Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab |
title | Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab |
title_full | Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab |
title_fullStr | Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab |
title_full_unstemmed | Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab |
title_short | Targeting Drosophila Sas6 to mitochondria reveals its high affinity for Gorab |
title_sort | targeting drosophila sas6 to mitochondria reveals its high affinity for gorab |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9836085/ https://www.ncbi.nlm.nih.gov/pubmed/36331102 http://dx.doi.org/10.1242/bio.059545 |
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