Molecular Components of Nitrate and Nitrite Efflux in Yeast

Cabrera, Elisa; González-Montelongo, Rafaela; Giráldez, Teresa; Rosa, Diego Álvarez de la; Siverio, José M.

doi:10.1128/ec.00268-13

Cited by 20 publications

(22 citation statements)

References 49 publications

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“…However, the aerial mycelium appeared to be thinner, suggesting less biomass formation ( Figure 1 ). This was confirmed by cultivation of the Wt in liquid medium with glutamine, nitrate or different concentrations of nitrite (Supplementary Figure S1B) and is probably caused by toxic effects of nitrite that inhibit growth of the fungus (Wodzinski et al, 1978; Crawford and Arst, 1993; Cabrera et al, 2014). The Δ NIAD and Δ NRTA mutants showed Wt-like growth on nitrite, whereas the Δ AREB mutant grew with a slightly reduced colony diameter compared to the Wt ( Figure 1 ).…”

Section: Resultsmentioning

confidence: 68%

Nitrate Assimilation in Fusarium fujikuroi Is Controlled by Multiple Levels of Regulation

2017

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Secondary metabolite production of the phytopathogenic ascomycete fungus Fusarium fujikuroi is greatly influenced by the availability of nitrogen. While favored nitrogen sources such as glutamine and ammonium are used preferentially, the uptake and utilization of nitrate is subject to a regulatory mechanism called nitrogen metabolite repression (NMR). In Aspergillus nidulans, the transcriptional control of the nitrate assimilatory system is carried out by the synergistic action of the nitrate-specific transcription factor NirA and the major nitrogen-responsive regulator AreA. In this study, we identified the main components of the nitrate assimilation system in F. fujikuroi and studied the role of each of them regarding the regulation of the remaining components. We analyzed mutants with deletions of the nitrate-specific activator NirA, the nitrate reductase (NR), the nitrite reductase (NiR) and the nitrate transporter NrtA. We show that NirA controls the transcription of the nitrate assimilatory genes NIAD, NIIA, and NRTA in the presence of nitrate, and that the global nitrogen regulator AreA is obligatory for expression of most, but not all NirA target genes (NIAD). By transforming a NirA-GFP fusion construct into the ΔNIAD, ΔNRTA, and ΔAREA mutant backgrounds we revealed that NirA was dispersed in the cytosol when grown in the presence of glutamine, but rapidly sorted to the nucleus when nitrate was added. Interestingly, the rapid and nitrate-induced nuclear translocation of NirA was observed also in the ΔAREA and ΔNRTA mutants, but not in ΔNIAD, suggesting that the fungus is able to directly sense nitrate in an AreA- and NrtA-independent, but NR-dependent manner.

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Section: Resultsmentioning

confidence: 68%

Nitrate Assimilation in Fusarium fujikuroi Is Controlled by Multiple Levels of Regulation

2017

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“…We were particularly interested in one gene required for grape juice survival in the lab strain . SSU1 is a plasma membrane sulfite/nitrate pump ( Park and Bakalinsky 2000 ; Sarver and DeRisi 2005 ; Mendes-Ferreira et al 2010 ; Cabrera et al 2014 ) that falls under the large-effect QTL on Chromosome XVI (Chr 16) in Cross 1. This gene has been linked to increased sulfite resistance ( Goto-Yamamoto et al 1998 ; Park and Bakalinsky 2000 ) as well as increased wine fermentation rates ( Yuasa et al 2005 ; Nardi et al 2010 ; Brion et al 2013 ; Zimmer et al 2014 ) and sulfur assimilation (a phenotype important to wine-yeast domestication) ( Mendes-Ferreira et al 2010 ).…”

Section: Resultsmentioning

confidence: 99%

Ecological and Genetic Barriers Differentiate Natural Populations ofSaccharomyces cerevisiae

et al. 2015

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How populations that inhabit the same geographical area become genetically differentiated is not clear. To investigate this, we characterized phenotypic and genetic differences between two populations of Saccharomyces cerevisiae that in some cases inhabit the same environment but show relatively little gene flow. We profiled stress sensitivity in a group of vineyard isolates and a group of oak-soil strains and found several niche-related phenotypes that distinguish the populations. We performed bulk-segregant mapping on two of the distinguishing traits: The vineyard-specific ability to grow in grape juice and oak-specific tolerance to the cell wall damaging drug Congo red. To implicate causal genes, we also performed a chemical genomic screen in the lab-strain deletion collection and identified many important genes that fell under quantitative trait loci peaks. One gene important for growth in grape juice and identified by both the mapping and the screen was SSU1, a sulfite-nitrite pump implicated in wine fermentations. The beneficial allele is generated by a known translocation that we reasoned may also serve as a genetic barrier. We found that the translocation is prevalent in vineyard strains, but absent in oak strains, and presents a postzygotic barrier to spore viability. Furthermore, the translocation was associated with a fitness cost to the rapid growth rate seen in oak-soil strains. Our results reveal the translocation as a dual-function locus that enforces ecological differentiation while producing a genetic barrier to gene flow in these sympatric populations.

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“…Interestingly, both SSU1 and FZF1 are also involved in the response to nitrosative stress (Sarver & DeRisi ; Cabrera et al . ), but the impact of this system in wild yeast populations is unknown.…”

Section: Discussionmentioning

confidence: 99%

Adaptive divergence in wine yeasts and their wild relatives suggests a prominent role for introgressions and rapid evolution at noncoding sites

et al. 2017

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In Saccharomyces cerevisiae, the main yeast in wine fermentation, the opportunity to examine divergence at the molecular level between a domesticated lineage and its wild counterpart arose recently due to the identification of the closest relatives of wine strains, a wild population associated with Mediterranean oaks. As genomic data are available for a considerable number of representatives belonging to both groups, we used population genomics to estimate the degree and distribution of nucleotide variation between wine yeasts and their closest wild relatives. We found widespread genomewide divergence, particularly at noncoding sites, which, together with above average divergence in trans-acting DNA binding proteins, may suggest an important role for divergence at the level of transcriptional regulation. Nine outlier regions putatively under strong divergent selection were highlighted by a genomewide scan under stringent conditions. Several cases of introgressions, originating in the sibling species Saccharomyces paradoxus, were also identified in the Mediterranean oak population. FZF1 and SSU1, mostly known for conferring sulphite resistance in wine yeasts, were among the introgressed genes, although not fixed. Because the introgressions detected in our study are not found in wine strains, we hypothesize that ongoing divergent ecological selection segregates the two forms between the different niches. Together, our results provide a first insight into the extent and kind of divergence between wine yeasts and their closest wild relatives.

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Molecular Components of Nitrate and Nitrite Efflux in Yeast

Cited by 20 publications

References 49 publications

Nitrate Assimilation in Fusarium fujikuroi Is Controlled by Multiple Levels of Regulation

Nitrate Assimilation in Fusarium fujikuroi Is Controlled by Multiple Levels of Regulation

Ecological and Genetic Barriers Differentiate Natural Populations ofSaccharomyces cerevisiae

Adaptive divergence in wine yeasts and their wild relatives suggests a prominent role for introgressions and rapid evolution at noncoding sites

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