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Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation
Studies of synthetic, well-defined biomolecular systems can elucidate inherent capabilities that may be difficult to uncover in a native biological context. Here, we used a minimal, reconstituted translation system from Escherichia coli to identify efficient ribosome binding sites (RBSs) in an unbia...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3310771/ https://www.ncbi.nlm.nih.gov/pubmed/22457640 http://dx.doi.org/10.1371/journal.pgen.1002598 |
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author | Barendt, Pamela A. Shah, Najaf A. Barendt, Gregory A. Sarkar, Casim A. |
author_facet | Barendt, Pamela A. Shah, Najaf A. Barendt, Gregory A. Sarkar, Casim A. |
author_sort | Barendt, Pamela A. |
collection | PubMed |
description | Studies of synthetic, well-defined biomolecular systems can elucidate inherent capabilities that may be difficult to uncover in a native biological context. Here, we used a minimal, reconstituted translation system from Escherichia coli to identify efficient ribosome binding sites (RBSs) in an unbiased, high-throughput manner. We applied ribosome display, a powerful in vitro selection method, to enrich only those mRNA sequences which could direct rapid protein translation. In addition to canonical Shine-Dalgarno (SD) motifs, we unexpectedly recovered highly efficient cytosine-rich (C-rich) sequences that exhibit unmistakable complementarity to the 16S rRNA of the small subunit of the ribosome, indicating that broad-specificity base-pairing may be an inherent, general mechanism for efficient translation. Furthermore, given the conservation of ribosomal structure and function across species, the broader relevance of C-rich RBS sequences identified through our in vitro evolution approach is supported by multiple, diverse examples in nature, including C-rich RBSs in several bacteriophage and plants, a poly-C consensus before the start codon in a lower eukaryote, and Kozak-like sequences in vertebrates. |
format | Online Article Text |
id | pubmed-3310771 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33107712012-03-28 Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation Barendt, Pamela A. Shah, Najaf A. Barendt, Gregory A. Sarkar, Casim A. PLoS Genet Research Article Studies of synthetic, well-defined biomolecular systems can elucidate inherent capabilities that may be difficult to uncover in a native biological context. Here, we used a minimal, reconstituted translation system from Escherichia coli to identify efficient ribosome binding sites (RBSs) in an unbiased, high-throughput manner. We applied ribosome display, a powerful in vitro selection method, to enrich only those mRNA sequences which could direct rapid protein translation. In addition to canonical Shine-Dalgarno (SD) motifs, we unexpectedly recovered highly efficient cytosine-rich (C-rich) sequences that exhibit unmistakable complementarity to the 16S rRNA of the small subunit of the ribosome, indicating that broad-specificity base-pairing may be an inherent, general mechanism for efficient translation. Furthermore, given the conservation of ribosomal structure and function across species, the broader relevance of C-rich RBS sequences identified through our in vitro evolution approach is supported by multiple, diverse examples in nature, including C-rich RBSs in several bacteriophage and plants, a poly-C consensus before the start codon in a lower eukaryote, and Kozak-like sequences in vertebrates. Public Library of Science 2012-03-22 /pmc/articles/PMC3310771/ /pubmed/22457640 http://dx.doi.org/10.1371/journal.pgen.1002598 Text en Barendt et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Barendt, Pamela A. Shah, Najaf A. Barendt, Gregory A. Sarkar, Casim A. Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation |
title | Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation |
title_full | Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation |
title_fullStr | Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation |
title_full_unstemmed | Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation |
title_short | Broad-Specificity mRNA–rRNA Complementarity in Efficient Protein Translation |
title_sort | broad-specificity mrna–rrna complementarity in efficient protein translation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3310771/ https://www.ncbi.nlm.nih.gov/pubmed/22457640 http://dx.doi.org/10.1371/journal.pgen.1002598 |
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