Cargando…
An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression
Riboregulators such as riboswitches and RNA thermometers provide simple, protein-independent tools to control gene expression at the post-transcriptional level. In bacteria, RNA thermometers regulate protein synthesis in response to temperature shifts. Thermometers outside of the bacterial world are...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10639063/ https://www.ncbi.nlm.nih.gov/pubmed/37855670 http://dx.doi.org/10.1093/nar/gkad816 |
_version_ | 1785146616951341056 |
---|---|
author | Chung, Kin Pan Loiacono, F Vanessa Neupert, Juliane Wu, Mengting Bock, Ralph |
author_facet | Chung, Kin Pan Loiacono, F Vanessa Neupert, Juliane Wu, Mengting Bock, Ralph |
author_sort | Chung, Kin Pan |
collection | PubMed |
description | Riboregulators such as riboswitches and RNA thermometers provide simple, protein-independent tools to control gene expression at the post-transcriptional level. In bacteria, RNA thermometers regulate protein synthesis in response to temperature shifts. Thermometers outside of the bacterial world are rare, and in organellar genomes, no RNA thermometers have been identified to date. Here we report the discovery of an RNA thermometer in a chloroplast gene of the unicellular green alga Chlamydomonas reinhardtii. The thermometer, residing in the 5′ untranslated region of the psaA messenger RNA forms a hairpin-type secondary structure that masks the Shine–Dalgarno sequence at 25°C. At 40°C, melting of the secondary structure increases accessibility of the Shine–Dalgarno sequence to initiating ribosomes, thus enhancing protein synthesis. By targeted nucleotide substitutions and transfer of the thermometer into Escherichia coli, we show that the secondary structure is necessary and sufficient to confer the thermometer properties. We also demonstrate that the thermometer provides a valuable tool for inducible transgene expression from the Chlamydomonas plastid genome, in that a simple temperature shift of the algal culture can greatly increase recombinant protein yields. |
format | Online Article Text |
id | pubmed-10639063 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-106390632023-11-15 An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression Chung, Kin Pan Loiacono, F Vanessa Neupert, Juliane Wu, Mengting Bock, Ralph Nucleic Acids Res Synthetic Biology and Bioengineering Riboregulators such as riboswitches and RNA thermometers provide simple, protein-independent tools to control gene expression at the post-transcriptional level. In bacteria, RNA thermometers regulate protein synthesis in response to temperature shifts. Thermometers outside of the bacterial world are rare, and in organellar genomes, no RNA thermometers have been identified to date. Here we report the discovery of an RNA thermometer in a chloroplast gene of the unicellular green alga Chlamydomonas reinhardtii. The thermometer, residing in the 5′ untranslated region of the psaA messenger RNA forms a hairpin-type secondary structure that masks the Shine–Dalgarno sequence at 25°C. At 40°C, melting of the secondary structure increases accessibility of the Shine–Dalgarno sequence to initiating ribosomes, thus enhancing protein synthesis. By targeted nucleotide substitutions and transfer of the thermometer into Escherichia coli, we show that the secondary structure is necessary and sufficient to confer the thermometer properties. We also demonstrate that the thermometer provides a valuable tool for inducible transgene expression from the Chlamydomonas plastid genome, in that a simple temperature shift of the algal culture can greatly increase recombinant protein yields. Oxford University Press 2023-10-19 /pmc/articles/PMC10639063/ /pubmed/37855670 http://dx.doi.org/10.1093/nar/gkad816 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. 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 | Synthetic Biology and Bioengineering Chung, Kin Pan Loiacono, F Vanessa Neupert, Juliane Wu, Mengting Bock, Ralph An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression |
title | An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression |
title_full | An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression |
title_fullStr | An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression |
title_full_unstemmed | An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression |
title_short | An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression |
title_sort | rna thermometer in the chloroplast genome of chlamydomonas facilitates temperature-controlled gene expression |
topic | Synthetic Biology and Bioengineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10639063/ https://www.ncbi.nlm.nih.gov/pubmed/37855670 http://dx.doi.org/10.1093/nar/gkad816 |
work_keys_str_mv | AT chungkinpan anrnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT loiaconofvanessa anrnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT neupertjuliane anrnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT wumengting anrnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT bockralph anrnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT chungkinpan rnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT loiaconofvanessa rnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT neupertjuliane rnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT wumengting rnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression AT bockralph rnathermometerinthechloroplastgenomeofchlamydomonasfacilitatestemperaturecontrolledgeneexpression |