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Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria

Endosymbiosis was fundamental for the evolution of eukaryotic complexity. Endosymbiotic interactions can be dissected through forward- and reverse-genetic experiments, such as RNA-interference (RNAi). However, distinguishing small (s)RNA pathways in a eukaryote–eukaryote endosymbiotic interaction is...

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Autores principales: Jenkins, Benjamin H., Maguire, Finlay, Leonard, Guy, Eaton, Joshua D., West, Steven, Housden, Benjamin E., Milner, David S., Richards, Thomas A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8059543/
https://www.ncbi.nlm.nih.gov/pubmed/33996132
http://dx.doi.org/10.1098/rsos.210140
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author Jenkins, Benjamin H.
Maguire, Finlay
Leonard, Guy
Eaton, Joshua D.
West, Steven
Housden, Benjamin E.
Milner, David S.
Richards, Thomas A.
author_facet Jenkins, Benjamin H.
Maguire, Finlay
Leonard, Guy
Eaton, Joshua D.
West, Steven
Housden, Benjamin E.
Milner, David S.
Richards, Thomas A.
author_sort Jenkins, Benjamin H.
collection PubMed
description Endosymbiosis was fundamental for the evolution of eukaryotic complexity. Endosymbiotic interactions can be dissected through forward- and reverse-genetic experiments, such as RNA-interference (RNAi). However, distinguishing small (s)RNA pathways in a eukaryote–eukaryote endosymbiotic interaction is challenging. Here, we investigate the repertoire of RNAi pathway protein-encoding genes in the model nascent endosymbiotic system, Paramecium bursaria–Chlorella spp. Using comparative genomics and transcriptomics supported by phylogenetics, we identify essential proteome components of the small interfering (si)RNA, scan (scn)RNA and internal eliminated sequence (ies)RNA pathways. Our analyses reveal that copies of these components have been retained throughout successive whole genome duplication (WGD) events in the Paramecium clade. We validate feeding-induced siRNA-based RNAi in P. bursaria via knock-down of the splicing factor, u2af1, which we show to be crucial to host growth. Finally, using simultaneous knock-down ‘paradox’ controls to rescue the effect of u2af1 knock-down, we demonstrate that feeding-induced RNAi in P. bursaria is dependent upon a core pathway of host-encoded Dcr1, Piwi and Pds1 components. Our experiments confirm the presence of a functional, host-derived RNAi pathway in P. bursaria that generates 23-nt siRNA, validating the use of the P. bursaria–Chlorella spp. system to investigate the genetic basis of a nascent endosymbiosis.
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spelling pubmed-80595432021-05-14 Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria Jenkins, Benjamin H. Maguire, Finlay Leonard, Guy Eaton, Joshua D. West, Steven Housden, Benjamin E. Milner, David S. Richards, Thomas A. R Soc Open Sci Biochemistry, Cellular and Molecular Biology Endosymbiosis was fundamental for the evolution of eukaryotic complexity. Endosymbiotic interactions can be dissected through forward- and reverse-genetic experiments, such as RNA-interference (RNAi). However, distinguishing small (s)RNA pathways in a eukaryote–eukaryote endosymbiotic interaction is challenging. Here, we investigate the repertoire of RNAi pathway protein-encoding genes in the model nascent endosymbiotic system, Paramecium bursaria–Chlorella spp. Using comparative genomics and transcriptomics supported by phylogenetics, we identify essential proteome components of the small interfering (si)RNA, scan (scn)RNA and internal eliminated sequence (ies)RNA pathways. Our analyses reveal that copies of these components have been retained throughout successive whole genome duplication (WGD) events in the Paramecium clade. We validate feeding-induced siRNA-based RNAi in P. bursaria via knock-down of the splicing factor, u2af1, which we show to be crucial to host growth. Finally, using simultaneous knock-down ‘paradox’ controls to rescue the effect of u2af1 knock-down, we demonstrate that feeding-induced RNAi in P. bursaria is dependent upon a core pathway of host-encoded Dcr1, Piwi and Pds1 components. Our experiments confirm the presence of a functional, host-derived RNAi pathway in P. bursaria that generates 23-nt siRNA, validating the use of the P. bursaria–Chlorella spp. system to investigate the genetic basis of a nascent endosymbiosis. The Royal Society 2021-04-21 /pmc/articles/PMC8059543/ /pubmed/33996132 http://dx.doi.org/10.1098/rsos.210140 Text en © 2021 The Authors. https://creativecommons.org/licenses/by/4.0/Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, provided the original author and source are credited.
spellingShingle Biochemistry, Cellular and Molecular Biology
Jenkins, Benjamin H.
Maguire, Finlay
Leonard, Guy
Eaton, Joshua D.
West, Steven
Housden, Benjamin E.
Milner, David S.
Richards, Thomas A.
Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria
title Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria
title_full Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria
title_fullStr Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria
title_full_unstemmed Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria
title_short Characterization of the RNA-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, Paramecium bursaria
title_sort characterization of the rna-interference pathway as a tool for reverse genetic analysis in the nascent phototrophic endosymbiosis, paramecium bursaria
topic Biochemistry, Cellular and Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8059543/
https://www.ncbi.nlm.nih.gov/pubmed/33996132
http://dx.doi.org/10.1098/rsos.210140
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