Cation/H+ antiporters mediate potassium and sodium fluxes in Pichia sorbitophila. Cloning of the PsNHA1 and PsNHA2 genes and expression in Saccharomyces cerevisiae

Bañuelos, María Antonia; Ramos, José; Calero, Fernando Sánchez; Braun, Valerie; Potier, Serge

doi:10.1002/yea.922

Cited by 23 publications

(25 citation statements)

References 31 publications

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“…The Escherichia coli strain DH5␣ was routinely used for plasmidic DNA propagation. The Saccharomyces cerevisiae strains used were W303.1A (Mat a ade2 ura3 trp1 leu2 his3) and its derivatives W⌬3 (Mat a ade2 ura3 trp1 trk1⌬::LEU2 trk2⌬::HIS3), which is deficient in the endogenous K ϩ uptake systems TRK1 and TRK2 (25), and MA5 (Mat a ade2 ura3 trp1 trk1⌬::LEU2 trk2⌬::HIS3 ena1-4⌬::HIS3 nha1⌬::LEU2), a strain obtained for this work by crossing the aforementioned W⌬3 with B31 (Mat a ade2 ura3 trp1 ena1-4⌬::HIS3 nha1⌬::LEU2) (6), which is deficient in the Na ϩ efflux systems ENA1-4 and NHA1. Fungal strains were normally grown either in the complex medium YPD (1% yeast extract, 2% peptone, 2% glucose) or in the minimal SD medium (48), supplemented with 50 mM K ϩ when used for the K ϩ uptake mutants.…”

Section: Methodsmentioning

confidence: 99%

Novel P-Type ATPases Mediate High-Affinity Potassium or Sodium Uptake in Fungi

et al. 2004

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Fungi have an absolute requirement for K؉ , but K ؉ may be partially replaced by Na ؉ . Na ؉ uptake in Ustilago maydis and Pichia sorbitophila was found to exhibit a fast rate, low K m , and apparent independence of the membrane potential. Searches of sequences with similarity to P-type ATPases in databases allowed us to identify three genes in these species, Umacu1, Umacu2, and PsACU1, that could encode P-type ATPases of a novel type. Deletion of the acu1 and acu2 genes proved that they encoded the transporters that mediated the high-affinity Na ؉ uptake of U. maydis. Heterologous expressions of the Umacu2 gene in K ؉ transport mutants of Saccharomyces cerevisiae and transport studies in the single and double ⌬acu1 and ⌬acu2 mutants of U. maydis revealed that the acu1 and acu2 genes encode transporters that mediated high-affinity K ؉ uptake in addition to Na ؉ uptake. Other fungi also have genes or pseudogenes whose translated sequences show high similarity to the ACU proteins of U. maydis and P. sorbitophila. In the phylogenetic tree of P-type ATPases all the identified ACU ATPases define a new cluster, which shows the lowest divergence with type IIC, animal Na ؉ ,K ؉ -ATPases. The fungal high-affinity Na ؉ uptake mediated by ACU ATPases is functionally identical to the uptake that is mediated by some plant HKT transporters. Potassium and Naϩ were present in the sea, where the early evolution of life took place, but only K ϩ was selected to maintain the electrical and osmotic equilibria of the cells. Constricted by the K ϩ environment, further evolution of the cellular physiology added to the original requirements; the K ϩ dependence on many physiologic processes that adapted to the cation was present. This K ϩ dependence of living cells was not a restriction for life in the sea, where K ϩ was abundant, but involved a nutritional problem for the start of terrestrial life on the rocks emerging from the sea, probably in the Precambrian (27). Therefore, before plants and fungi could colonize terrestrial environments (12, 26) they had to sustain a complex process of physiological adaptations to a new medium in which K ϩ and Na ϩ concentrations were very low. A crucial role in this terrestrial adaptation was played by high-affinity K ϩ transporters, whose existence has been well established for several plant and fungal species (44); but very little is known of other cooperative processes that may alleviate these strict K ϩ requirements. Among these, an emerging notion is that Na ϩ can be used as a substitute for K ϩ . It has been known for a long time that Na ϩ enhances the growth of fungi (17) and plants (14,28,49) under conditions of K ϩ deficiency, but this effect had been considered accidental or under a weak physiological regulation. In contrast with this view, the characterization of high-affinity Na ϩ uptake in several plants and the fact that it takes place only when K ϩ has been exhausted (21) strongly suggest that Na ϩ plays a physiologically programmed role in plants given an insufficient supply of K ϩ ....

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Section: Methodsmentioning

confidence: 99%

Novel P-Type ATPases Mediate High-Affinity Potassium or Sodium Uptake in Fungi

et al. 2004

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“…The genomes of most yeast species (e.g. S. cerevisiae, Candida albicans, Pichia sorbithophila, Debaryomyces hansenii) contain only one gene encoding a Na + /H + antiporter with a broad substrate specificity for Na + , Li + , K + and Rb + (Banuelos et al, 1998(Banuelos et al, , 2002Kinclova et al, 2001b, c;Velkova & Sychrová, 2006). Only two species (Yarrowia lipolytica, Schizosaccharomyces pombe) possess two types of plasma-membrane Nha1/ Sod2 antiporter: one of them primarily transports Na + (Li + ) cations, the other mainly transports K + (and Rb + ) (Papouskova & Sychrová, 2006, 2007.…”

Section: Introductionmentioning

confidence: 99%

Plasma-membrane Cnh1 Na+/H+ antiporter regulates potassium homeostasis in Candida albicans

Zimmermannová

Sychrová

2007

Microbiology

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“…Besides S. cerevisiae, it was identified in the osmotolerant D. hansenii (273), in P. sorbitophila (14), and in five members of the Candida family, i.e., C. albicans, C. dubliniensis, C. parapsilosis, C. glabrata, and C. tropicalis (113,117,131). All of these transporters have been characterized by heterologous expression in S. cerevisiae cells lacking the NHA1 and ENA1-4 genes, i.e., in cells unable to export alkali metal cations efficiently.…”

Section: Namentioning

confidence: 99%

Alkali Metal Cation Transport and Homeostasis in Yeasts

Ariño

Ramos

Sychrová

2010

Microbiol Mol Biol Rev

261

299

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SUMMARY The maintenance of appropriate intracellular concentrations of alkali metal cations, principally K+ and Na+, is of utmost importance for living cells, since they determine cell volume, intracellular pH, and potential across the plasma membrane, among other important cellular parameters. Yeasts have developed a number of strategies to adapt to large variations in the concentrations of these cations in the environment, basically by controlling transport processes. Plasma membrane high-affinity K+ transporters allow intracellular accumulation of this cation even when it is scarce in the environment. Exposure to high concentrations of Na+ can be tolerated due to the existence of an Na+, K+-ATPase and an Na+, K+/H+-antiporter, which contribute to the potassium balance as well. Cations can also be sequestered through various antiporters into intracellular organelles, such as the vacuole. Although some uncertainties still persist, the nature of the major structural components responsible for alkali metal cation fluxes across yeast membranes has been defined within the last 20 years. In contrast, the regulatory components and their interactions are, in many cases, still unclear. Conserved signaling pathways (e.g., calcineurin and HOG) are known to participate in the regulation of influx and efflux processes at the plasma membrane level, even though the molecular details are obscure. Similarly, very little is known about the regulation of organellar transport and homeostasis of alkali metal cations. The aim of this review is to provide a comprehensive and up-to-date vision of the mechanisms responsible for alkali metal cation transport and their regulation in the model yeast Saccharomyces cerevisiae and to establish, when possible, comparisons with other yeasts and higher plants.

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Cation/H⁺ antiporters mediate potassium and sodium fluxes in Pichia sorbitophila. Cloning of the PsNHA1 and PsNHA2 genes and expression in Saccharomyces cerevisiae

Cited by 23 publications

References 31 publications

Novel P-Type ATPases Mediate High-Affinity Potassium or Sodium Uptake in Fungi

Novel P-Type ATPases Mediate High-Affinity Potassium or Sodium Uptake in Fungi

Plasma-membrane Cnh1 Na+/H+ antiporter regulates potassium homeostasis in Candida albicans

Alkali Metal Cation Transport and Homeostasis in Yeasts

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