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Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection
Plastids are the site of complex biochemical pathways, most prominently photosynthesis. The organelle evolved through endosymbiosis with a cyanobacterium, which is exemplified by the outer envelope membrane that harbors more than 40 proteins in Arabidopsis. Their evolutionary conservation indicates...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10542568/ https://www.ncbi.nlm.nih.gov/pubmed/37572358 http://dx.doi.org/10.1093/g3journal/jkad187 |
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author | Schwenkert, Serena Lo, Wing Tung Szulc, Beata Yip, Chun Kwan Pratt, Anna I Cusack, Siobhan A Brandt, Benjamin Leister, Dario Kunz, Hans-Henning |
author_facet | Schwenkert, Serena Lo, Wing Tung Szulc, Beata Yip, Chun Kwan Pratt, Anna I Cusack, Siobhan A Brandt, Benjamin Leister, Dario Kunz, Hans-Henning |
author_sort | Schwenkert, Serena |
collection | PubMed |
description | Plastids are the site of complex biochemical pathways, most prominently photosynthesis. The organelle evolved through endosymbiosis with a cyanobacterium, which is exemplified by the outer envelope membrane that harbors more than 40 proteins in Arabidopsis. Their evolutionary conservation indicates high significance for plant cell function. While a few proteins are well-studied as part of the protein translocon complex the majority of outer envelope protein functions is unclear. Gaining a deeper functional understanding has been complicated by the lack of observable loss-of-function mutant phenotypes, which is often rooted in functional genetic redundancy. Therefore, we designed outer envelope-specific artificial micro RNAs (oemiRs) capable of downregulating transcripts from several loci simultaneously. We successfully tested oemiR function by performing a proof-of-concept screen for pale and cold-sensitive mutants. An in-depth analysis of pale mutant alleles deficient in the translocon component TOC75 using proteomics provided new insights into putative compensatory import pathways. The cold stress screen not only recapitulated 3 previously known phenotypes of cold-sensitive mutants but also identified 4 mutants of additional oemiR outer envelope loci. Altogether our study revealed a role of the outer envelope to tolerate cold conditions and showcasts the power of the oemiR collection to research the significance of outer envelope proteins. |
format | Online Article Text |
id | pubmed-10542568 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-105425682023-10-03 Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection Schwenkert, Serena Lo, Wing Tung Szulc, Beata Yip, Chun Kwan Pratt, Anna I Cusack, Siobhan A Brandt, Benjamin Leister, Dario Kunz, Hans-Henning G3 (Bethesda) Mutant Screen Report Plastids are the site of complex biochemical pathways, most prominently photosynthesis. The organelle evolved through endosymbiosis with a cyanobacterium, which is exemplified by the outer envelope membrane that harbors more than 40 proteins in Arabidopsis. Their evolutionary conservation indicates high significance for plant cell function. While a few proteins are well-studied as part of the protein translocon complex the majority of outer envelope protein functions is unclear. Gaining a deeper functional understanding has been complicated by the lack of observable loss-of-function mutant phenotypes, which is often rooted in functional genetic redundancy. Therefore, we designed outer envelope-specific artificial micro RNAs (oemiRs) capable of downregulating transcripts from several loci simultaneously. We successfully tested oemiR function by performing a proof-of-concept screen for pale and cold-sensitive mutants. An in-depth analysis of pale mutant alleles deficient in the translocon component TOC75 using proteomics provided new insights into putative compensatory import pathways. The cold stress screen not only recapitulated 3 previously known phenotypes of cold-sensitive mutants but also identified 4 mutants of additional oemiR outer envelope loci. Altogether our study revealed a role of the outer envelope to tolerate cold conditions and showcasts the power of the oemiR collection to research the significance of outer envelope proteins. Oxford University Press 2023-08-12 /pmc/articles/PMC10542568/ /pubmed/37572358 http://dx.doi.org/10.1093/g3journal/jkad187 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of The Genetics Society of America. 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 reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Mutant Screen Report Schwenkert, Serena Lo, Wing Tung Szulc, Beata Yip, Chun Kwan Pratt, Anna I Cusack, Siobhan A Brandt, Benjamin Leister, Dario Kunz, Hans-Henning Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection |
title | Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection |
title_full | Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection |
title_fullStr | Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection |
title_full_unstemmed | Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection |
title_short | Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection |
title_sort | probing the physiological role of the plastid outer-envelope membrane using the oemir plasmid collection |
topic | Mutant Screen Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10542568/ https://www.ncbi.nlm.nih.gov/pubmed/37572358 http://dx.doi.org/10.1093/g3journal/jkad187 |
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