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Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus

It is important to understand the basis of thermotolerance in yeasts to broaden their application in industrial biotechnology. The capacity to run bioprocesses at temperatures above 40 °C is of great interest but this is beyond the growth range of most of the commonly used yeast species. In contrast...

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Autores principales: Montini, Noemi, Doughty, Tyler W., Domenzain, Iván, Fenton, Darren A., Baranov, Pavel V., Harrington, Ronan, Nielsen, Jens, Siewers, Verena, Morrissey, John P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Microbiology Society 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9558357/
https://www.ncbi.nlm.nih.gov/pubmed/35333706
http://dx.doi.org/10.1099/mic.0.001148
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author Montini, Noemi
Doughty, Tyler W.
Domenzain, Iván
Fenton, Darren A.
Baranov, Pavel V.
Harrington, Ronan
Nielsen, Jens
Siewers, Verena
Morrissey, John P.
author_facet Montini, Noemi
Doughty, Tyler W.
Domenzain, Iván
Fenton, Darren A.
Baranov, Pavel V.
Harrington, Ronan
Nielsen, Jens
Siewers, Verena
Morrissey, John P.
author_sort Montini, Noemi
collection PubMed
description It is important to understand the basis of thermotolerance in yeasts to broaden their application in industrial biotechnology. The capacity to run bioprocesses at temperatures above 40 °C is of great interest but this is beyond the growth range of most of the commonly used yeast species. In contrast, some industrial yeasts such as Kluyveromyces marxianus can grow at temperatures of 45 °C or higher. Such species are valuable for direct use in industrial biotechnology and as a vehicle to study the genetic and physiological basis of yeast thermotolerance. In previous work, we reported that evolutionarily young genes disproportionately changed expression when yeast were growing under stressful conditions and postulated that such genes could be important for long-term adaptation to stress. Here, we tested this hypothesis in K. marxianus by identifying and studying species-specific genes that showed increased expression during high-temperature growth. Twelve such genes were identified and 11 were successfully inactivated using CRISPR-mediated mutagenesis. One gene, KLMX_70384, is required for competitive growth at high temperature, supporting the hypothesis that evolutionary young genes could play roles in adaptation to harsh environments. KLMX_70384 is predicted to encode an 83 aa peptide, and RNA sequencing and ribo-sequencing were used to confirm transcription and translation of the gene. The precise function of KLMX_70384 remains unknown but some features are suggestive of RNA-binding activity. The gene is located in what was previously considered an intergenic region of the genome, which lacks homologues in other yeasts or in databases. Overall, the data support the hypothesis that genes that arose de novo in K. marxianus after the speciation event that separated K. marxianus and K. lactis contribute to some of its unique traits.
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spelling pubmed-95583572022-10-14 Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus Montini, Noemi Doughty, Tyler W. Domenzain, Iván Fenton, Darren A. Baranov, Pavel V. Harrington, Ronan Nielsen, Jens Siewers, Verena Morrissey, John P. Microbiology (Reading) Microbial Physiology, Biochemistry and Metabolism (formerly Physiology and Metabolism) It is important to understand the basis of thermotolerance in yeasts to broaden their application in industrial biotechnology. The capacity to run bioprocesses at temperatures above 40 °C is of great interest but this is beyond the growth range of most of the commonly used yeast species. In contrast, some industrial yeasts such as Kluyveromyces marxianus can grow at temperatures of 45 °C or higher. Such species are valuable for direct use in industrial biotechnology and as a vehicle to study the genetic and physiological basis of yeast thermotolerance. In previous work, we reported that evolutionarily young genes disproportionately changed expression when yeast were growing under stressful conditions and postulated that such genes could be important for long-term adaptation to stress. Here, we tested this hypothesis in K. marxianus by identifying and studying species-specific genes that showed increased expression during high-temperature growth. Twelve such genes were identified and 11 were successfully inactivated using CRISPR-mediated mutagenesis. One gene, KLMX_70384, is required for competitive growth at high temperature, supporting the hypothesis that evolutionary young genes could play roles in adaptation to harsh environments. KLMX_70384 is predicted to encode an 83 aa peptide, and RNA sequencing and ribo-sequencing were used to confirm transcription and translation of the gene. The precise function of KLMX_70384 remains unknown but some features are suggestive of RNA-binding activity. The gene is located in what was previously considered an intergenic region of the genome, which lacks homologues in other yeasts or in databases. Overall, the data support the hypothesis that genes that arose de novo in K. marxianus after the speciation event that separated K. marxianus and K. lactis contribute to some of its unique traits. Microbiology Society 2022-03-25 /pmc/articles/PMC9558357/ /pubmed/35333706 http://dx.doi.org/10.1099/mic.0.001148 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
spellingShingle Microbial Physiology, Biochemistry and Metabolism (formerly Physiology and Metabolism)
Montini, Noemi
Doughty, Tyler W.
Domenzain, Iván
Fenton, Darren A.
Baranov, Pavel V.
Harrington, Ronan
Nielsen, Jens
Siewers, Verena
Morrissey, John P.
Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus
title Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus
title_full Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus
title_fullStr Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus
title_full_unstemmed Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus
title_short Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast Kluyveromyces marxianus
title_sort identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeast kluyveromyces marxianus
topic Microbial Physiology, Biochemistry and Metabolism (formerly Physiology and Metabolism)
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9558357/
https://www.ncbi.nlm.nih.gov/pubmed/35333706
http://dx.doi.org/10.1099/mic.0.001148
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