An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must
© 2014, Springer-Verlag Berlin Heidelberg. We used experimental evolution in order to identify genes involved in the adaptation of Saccharomyces cerevisiae to the early stages of alcoholic fermentation. Evolution experiments were run for about 200 generations, in continuous culture conditions emulating the initial stages of wine fermentation. We performed whole-genome sequencing of four adapted strains from three independent evolution experiments. Mutations identified in these strains pointed to the Rsp5p-Bul1/2p ubiquitin ligase complex as the preferred evolutionary target under these experimental conditions. Rsp5p is a multifunctional enzyme able to ubiquitinate target proteins participating in different cellular processes, while Bul1p is an Rsp5p substrate adaptor specifically involved in the ubiquitin-dependent internalization of Gap1p and other plasma membrane permeases. While a loss-of-function mutation in BUL1 seems to be enough to confer a selective advantage under these assay conditions, this did not seem to be the case for RSP5 mutated strains, which required additional mutations, probably compensating for the detrimental effect of altered Rsp5p activity on essential cellular functions. The power of this experimental approach is illustrated by the identification of four independent mutants, each with a limited number of SNPs, affected within the same pathway. However, in order to obtain information relevant for a specific biotechnological process, caution must be taken in the choice of the background yeast genotype (as shown in this case for auxotrophies). In addition, the use of very stable continuous fermentation conditions might lead to the selection of a rather limited number of adaptive responses that would mask other possible targets for genetic improvement.
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Springer
2014-02
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Subjects: | Wine yeast, Next-generation sequencing (NGS), Fermentation kinetics, Experimental evolution, Adaptive laboratory evolution, Rsp5p-Bul1/2p, |
Online Access: | http://hdl.handle.net/10261/144150 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100004837 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100011011 |
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dig-icvv-es-10261-1441502020-05-21T06:48:30Z An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must Mangado, Ana Tronchoni, Jordi Morales, Pilar Novo, Maite Quirós Asensio, Manuel González García, Ramón Consejo Superior de Investigaciones Científicas (España) Ministerio de Economía y Competitividad (España) Junta de Andalucía Ministerio de Ciencia e Innovación (España) European Commission Wine yeast Next-generation sequencing (NGS) Fermentation kinetics Experimental evolution Adaptive laboratory evolution Rsp5p-Bul1/2p © 2014, Springer-Verlag Berlin Heidelberg. We used experimental evolution in order to identify genes involved in the adaptation of Saccharomyces cerevisiae to the early stages of alcoholic fermentation. Evolution experiments were run for about 200 generations, in continuous culture conditions emulating the initial stages of wine fermentation. We performed whole-genome sequencing of four adapted strains from three independent evolution experiments. Mutations identified in these strains pointed to the Rsp5p-Bul1/2p ubiquitin ligase complex as the preferred evolutionary target under these experimental conditions. Rsp5p is a multifunctional enzyme able to ubiquitinate target proteins participating in different cellular processes, while Bul1p is an Rsp5p substrate adaptor specifically involved in the ubiquitin-dependent internalization of Gap1p and other plasma membrane permeases. While a loss-of-function mutation in BUL1 seems to be enough to confer a selective advantage under these assay conditions, this did not seem to be the case for RSP5 mutated strains, which required additional mutations, probably compensating for the detrimental effect of altered Rsp5p activity on essential cellular functions. The power of this experimental approach is illustrated by the identification of four independent mutants, each with a limited number of SNPs, affected within the same pathway. However, in order to obtain information relevant for a specific biotechnological process, caution must be taken in the choice of the background yeast genotype (as shown in this case for auxotrophies). In addition, the use of very stable continuous fermentation conditions might lead to the selection of a rather limited number of adaptive responses that would mask other possible targets for genetic improvement. This work was supported by the Spanish Ministerio de Ciencia e Innovación (grants AGL2009-07327 and AGL2012-32064) and Junta de Andalucía (grant P10-AGR6544). AM was the recipient of a FPI fellowship from the Spanish Ministerio de Economía y Competitividad. MQ and MN were recipients of JAE-Doc fellowships from the Spanish National Research Council (CSIC), co-funded by the European Social Fund of the EU. Peer Reviewed 2017-02-17T08:45:11Z 2017-02-17T08:45:11Z 2014-02 2017-02-17T08:45:12Z artículo http://purl.org/coar/resource_type/c_6501 e-issn: 1432-0614 issn: 0175-7598 Applied Microbiology and Biotechnology 99(3): 1273-1286 (2014) http://hdl.handle.net/10261/144150 10.1007/s00253-014-6126-4 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100004837 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100011011 http://doi.org/10.1007/s00253-014-6126-4 Sí none Springer |
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Wine yeast Next-generation sequencing (NGS) Fermentation kinetics Experimental evolution Adaptive laboratory evolution Rsp5p-Bul1/2p Wine yeast Next-generation sequencing (NGS) Fermentation kinetics Experimental evolution Adaptive laboratory evolution Rsp5p-Bul1/2p |
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Wine yeast Next-generation sequencing (NGS) Fermentation kinetics Experimental evolution Adaptive laboratory evolution Rsp5p-Bul1/2p Wine yeast Next-generation sequencing (NGS) Fermentation kinetics Experimental evolution Adaptive laboratory evolution Rsp5p-Bul1/2p Mangado, Ana Tronchoni, Jordi Morales, Pilar Novo, Maite Quirós Asensio, Manuel González García, Ramón An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must |
description |
© 2014, Springer-Verlag Berlin Heidelberg. We used experimental evolution in order to identify genes involved in the adaptation of Saccharomyces cerevisiae to the early stages of alcoholic fermentation. Evolution experiments were run for about 200 generations, in continuous culture conditions emulating the initial stages of wine fermentation. We performed whole-genome sequencing of four adapted strains from three independent evolution experiments. Mutations identified in these strains pointed to the Rsp5p-Bul1/2p ubiquitin ligase complex as the preferred evolutionary target under these experimental conditions. Rsp5p is a multifunctional enzyme able to ubiquitinate target proteins participating in different cellular processes, while Bul1p is an Rsp5p substrate adaptor specifically involved in the ubiquitin-dependent internalization of Gap1p and other plasma membrane permeases. While a loss-of-function mutation in BUL1 seems to be enough to confer a selective advantage under these assay conditions, this did not seem to be the case for RSP5 mutated strains, which required additional mutations, probably compensating for the detrimental effect of altered Rsp5p activity on essential cellular functions. The power of this experimental approach is illustrated by the identification of four independent mutants, each with a limited number of SNPs, affected within the same pathway. However, in order to obtain information relevant for a specific biotechnological process, caution must be taken in the choice of the background yeast genotype (as shown in this case for auxotrophies). In addition, the use of very stable continuous fermentation conditions might lead to the selection of a rather limited number of adaptive responses that would mask other possible targets for genetic improvement. |
author2 |
Consejo Superior de Investigaciones Científicas (España) |
author_facet |
Consejo Superior de Investigaciones Científicas (España) Mangado, Ana Tronchoni, Jordi Morales, Pilar Novo, Maite Quirós Asensio, Manuel González García, Ramón |
format |
artículo |
topic_facet |
Wine yeast Next-generation sequencing (NGS) Fermentation kinetics Experimental evolution Adaptive laboratory evolution Rsp5p-Bul1/2p |
author |
Mangado, Ana Tronchoni, Jordi Morales, Pilar Novo, Maite Quirós Asensio, Manuel González García, Ramón |
author_sort |
Mangado, Ana |
title |
An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must |
title_short |
An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must |
title_full |
An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must |
title_fullStr |
An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must |
title_full_unstemmed |
An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must |
title_sort |
impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must |
publisher |
Springer |
publishDate |
2014-02 |
url |
http://hdl.handle.net/10261/144150 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100004837 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100011011 |
work_keys_str_mv |
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