Role of the DHH1 gene in the regulation of monocarboxylic acids transporters expression in Saccharomyces cerevisiae

Sandra Mota; Neide Vieira; Sónia Barbosa; Thierry Delaveau; Claire Torchet; Agnès Le Saux; Mathilde Garcia; Ana Pereira; Sophie Lemoine; Fanny Coulpier; Xavier Darzacq; Lionel Benard; Margarida Casal; Frédéric Devaux; Sandra Paiva

doi:10.1371/journal.pone.0111589

Role of the DHH1 gene in the regulation of monocarboxylic acids transporters expression in Saccharomyces cerevisiae

PLoS One. 2014 Nov 3;9(11):e111589. doi: 10.1371/journal.pone.0111589. eCollection 2014.

Authors

Affiliations

¹ Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Campus de Gualtar, Braga, Portugal; Centre of Health and Environmental Research (CISA), School of Allied Health Sciences, Polytechnic Institute of Porto, Vila Nova de Gaia, Portugal.
² Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Campus de Gualtar, Braga, Portugal.
³ Sorbonne Universités, Université Pierre et Marie Curie, UMR7238, Laboratoire de Biologie computationnelle et quantitative, Paris, France; CNRS, UMR7238, Laboratoire de Biologie computationnelle et quantitative, Paris, France.
⁴ CNRS, UMR8226, Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes, Institut de Biologie Physico-Chimique, Paris, France; Sorbonne Universités, Université Pierre et Marie Curie UPMC, UMR8226, Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes, Institut de Biologie Physico-Chimique, Paris, France.
⁵ CNRS, FRE3630, Laboratoire de l'Expression Génétique Microbienne, Institut de Biologie Physico-Chimique, Paris, France.
⁶ École normale supérieure, Institut de Biologie de l'ENS, IBENS, Paris, France; Inserm, U1024, Paris, France; CNRS, UMR 8197, Paris, France.

Abstract

Previous experiments revealed that DHH1, a RNA helicase involved in the regulation of mRNA stability and translation, complemented the phenotype of a Saccharomyces cerevisiae mutant affected in the expression of genes coding for monocarboxylic-acids transporters, JEN1 and ADY2 (Paiva S, Althoff S, Casal M, Leao C. FEMS Microbiol Lett, 1999, 170:301-306). In wild type cells, JEN1 expression had been shown to be undetectable in the presence of glucose or formic acid, and induced in the presence of lactate. In this work, we show that JEN1 mRNA accumulates in a dhh1 mutant, when formic acid was used as sole carbon source. Dhh1 interacts with the decapping activator Dcp1 and with the deadenylase complex. This led to the hypothesis that JEN1 expression is post-transcriptionally regulated by Dhh1 in formic acid. Analyses of JEN1 mRNAs decay in wild-type and dhh1 mutant strains confirmed this hypothesis. In these conditions, the stabilized JEN1 mRNA was associated to polysomes but no Jen1 protein could be detected, either by measurable lactate carrier activity, Jen1-GFP fluorescence detection or western blots. These results revealed the complexity of the expression regulation of JEN1 in S. cerevisiae and evidenced the importance of DHH1 in this process. Additionally, microarray analyses of dhh1 mutant indicated that Dhh1 plays a large role in metabolic adaptation, suggesting that carbon source changes triggers a complex interplay between transcriptional and post-transcriptional effects.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptation, Physiological
DEAD-box RNA Helicases / genetics*
DEAD-box RNA Helicases / metabolism*
Formates / metabolism
Gene Expression Profiling
Gene Expression Regulation, Fungal*
Genome-Wide Association Study
Monocarboxylic Acid Transporters / genetics*
Monocarboxylic Acid Transporters / metabolism
Mutation
Polyribosomes / metabolism
RNA Stability
RNA, Messenger / genetics
RNA, Messenger / metabolism
Saccharomyces cerevisiae / genetics*
Saccharomyces cerevisiae / growth & development
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / genetics*
Saccharomyces cerevisiae Proteins / metabolism*
Symporters / genetics
Symporters / metabolism

Substances

Formates
JEN1 protein, S cerevisiae
Monocarboxylic Acid Transporters
RNA, Messenger
Saccharomyces cerevisiae Proteins
Symporters
formic acid
DHH1 protein, S cerevisiae
DEAD-box RNA Helicases

Grants and funding

This study was supported by the Portuguese grant POCI/BIA-BCM/57812/2004 (Eixo 2, Medida 2.3, QCAIII – FEDER). N.V. received a FCT PhD fellowship (SFRH/BD/23503/2005). S.M. received a FCT PhD fellowship (SFRH/BD/74790/2010). F.D.’s work is supported by a grant from the Agence pour la Recherche contre le Cancer (ARC). Support to C.B.M.A. by FEDER through POFC-COMPETE and by Portuguese funds from FCT through the project PEst-OE/BIA/UI4050/2014 is also acknowledged. The authors thank Josette Banroques and Kyle Tanner for their advice regarding polysome gradients. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.