caa a22 4500 605501564 CHVBK 20210128100601.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00253-014-6126-4 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6126-4 An impaired ubiquitin ligase complex favors initial growth of auxotrophic yeast strains in synthetic grape must [Elektronische Daten] [Ana Mangado, Jordi Tronchoni, Pilar Morales, Maite Novo, Manuel Quirós, Ramon Gonzalez] 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. Springer-Verlag Berlin Heidelberg, 2014 Wine yeast nationallicence Adaptive laboratory evolution (ALE) nationallicence Experimental evolution nationallicence Fermentation kinetics nationallicence Next-generation sequencing (NGS) nationallicence Rsp5p-Bul1/2p nationallicence Mangado Ana Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut Tronchoni Jordi Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut Morales Pilar Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut Novo Maite Departament de Bioquímica i Biotecnologia, Facultat de Ciències de l′Educació i Psicologia, Universitat Rovira i Virgili, Tarragona, Spain aut Quirós Manuel Evolva Biotech A/S, Copenhagen Ø, Denmark aut Gonzalez Ramon Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/3(2015-02-01), 1273-1286 0175-7598 99:3<1273 2015 99 253 https://doi.org/10.1007/s00253-014-6126-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6126-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mangado Ana Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut NATIONALLICENCE 700 1- Tronchoni Jordi Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut NATIONALLICENCE 700 1- Morales Pilar Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut NATIONALLICENCE 700 1- Novo Maite Departament de Bioquímica i Biotecnologia, Facultat de Ciències de l′Educació i Psicologia, Universitat Rovira i Virgili, Tarragona, Spain aut NATIONALLICENCE 700 1- Quirós Manuel Evolva Biotech A/S, Copenhagen Ø, Denmark aut NATIONALLICENCE 700 1- Gonzalez Ramon Instituto de Ciencias de la Vid y del Vino, ICVV, (CSIC-Universidad de La Rioja-Gobierno de La Rioja), Madre de Dios 51, 26006, Logroño, La Rioja, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/3(2015-02-01), 1273-1286 0175-7598 99:3<1273 2015 99 253