Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae

Post-transcriptional upregulation is an effective way to increase the expression of transgenes and thus maximize the yields of target chemicals from metabolically engineered organisms. Refractory elements in the 3′ untranslated region (UTR) that increase mRNA half-life might be available. In Sacchar...

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Autores principales: Ito, Yoichiro, Kitagawa, Takao, Yamanishi, Mamoru, Katahira, Satoshi, Izawa, Shingo, Irie, Kenji, Furutani-Seiki, Makoto, Matsuyama, Takashi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5109538/
https://www.ncbi.nlm.nih.gov/pubmed/27845367
http://dx.doi.org/10.1038/srep36997
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author Ito, Yoichiro
Kitagawa, Takao
Yamanishi, Mamoru
Katahira, Satoshi
Izawa, Shingo
Irie, Kenji
Furutani-Seiki, Makoto
Matsuyama, Takashi
author_facet Ito, Yoichiro
Kitagawa, Takao
Yamanishi, Mamoru
Katahira, Satoshi
Izawa, Shingo
Irie, Kenji
Furutani-Seiki, Makoto
Matsuyama, Takashi
author_sort Ito, Yoichiro
collection PubMed
description Post-transcriptional upregulation is an effective way to increase the expression of transgenes and thus maximize the yields of target chemicals from metabolically engineered organisms. Refractory elements in the 3′ untranslated region (UTR) that increase mRNA half-life might be available. In Saccharomyces cerevisiae, several terminator regions have shown activity in increasing the production of proteins by upstream coding genes; among these terminators the DIT1 terminator has the highest activity. Here, we found in Saccharomyces cerevisiae that two resident trans-acting RNA-binding proteins (Nab6p and Pap1p) enhance the activity of the DIT1 terminator through the cis element GUUCG/U within the 3′-UTR. These two RNA-binding proteins could upregulate a battery of cell-wall–related genes. Mutagenesis of the DIT1 terminator improved its activity by a maximum of 500% of that of the standard PGK1 terminator. Further understanding and improvement of this system will facilitate inexpensive and stable production of complicated organism-derived drugs worldwide.
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spelling pubmed-51095382016-11-25 Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae Ito, Yoichiro Kitagawa, Takao Yamanishi, Mamoru Katahira, Satoshi Izawa, Shingo Irie, Kenji Furutani-Seiki, Makoto Matsuyama, Takashi Sci Rep Article Post-transcriptional upregulation is an effective way to increase the expression of transgenes and thus maximize the yields of target chemicals from metabolically engineered organisms. Refractory elements in the 3′ untranslated region (UTR) that increase mRNA half-life might be available. In Saccharomyces cerevisiae, several terminator regions have shown activity in increasing the production of proteins by upstream coding genes; among these terminators the DIT1 terminator has the highest activity. Here, we found in Saccharomyces cerevisiae that two resident trans-acting RNA-binding proteins (Nab6p and Pap1p) enhance the activity of the DIT1 terminator through the cis element GUUCG/U within the 3′-UTR. These two RNA-binding proteins could upregulate a battery of cell-wall–related genes. Mutagenesis of the DIT1 terminator improved its activity by a maximum of 500% of that of the standard PGK1 terminator. Further understanding and improvement of this system will facilitate inexpensive and stable production of complicated organism-derived drugs worldwide. Nature Publishing Group 2016-11-15 /pmc/articles/PMC5109538/ /pubmed/27845367 http://dx.doi.org/10.1038/srep36997 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Ito, Yoichiro
Kitagawa, Takao
Yamanishi, Mamoru
Katahira, Satoshi
Izawa, Shingo
Irie, Kenji
Furutani-Seiki, Makoto
Matsuyama, Takashi
Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae
title Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae
title_full Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae
title_fullStr Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae
title_full_unstemmed Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae
title_short Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae
title_sort enhancement of protein production via the strong dit1 terminator and two rna-binding proteins in saccharomyces cerevisiae
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5109538/
https://www.ncbi.nlm.nih.gov/pubmed/27845367
http://dx.doi.org/10.1038/srep36997
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