Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i>
Plasma membrane H<sup>+</sup>-ATPases of fungi, yeasts, and plants act as proton pumps to generate an electrochemical gradient, which is essential for secondary transport and intracellular pH maintenance. <i>Saccharomyces cerevisiae</i> has two genes (PMA1 and PMA2) encoding...
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MDPI AG
2022-05-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/8/6/550 |
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| author | Melissa Vázquez-Carrada Michael Feldbrügge Dario Rafael Olicón-Hernández Guadalupe Guerra-Sánchez Juan Pablo Pardo |
| author_facet | Melissa Vázquez-Carrada Michael Feldbrügge Dario Rafael Olicón-Hernández Guadalupe Guerra-Sánchez Juan Pablo Pardo |
| author_sort | Melissa Vázquez-Carrada |
| collection | DOAJ |
| description | Plasma membrane H<sup>+</sup>-ATPases of fungi, yeasts, and plants act as proton pumps to generate an electrochemical gradient, which is essential for secondary transport and intracellular pH maintenance. <i>Saccharomyces cerevisiae</i> has two genes (PMA1 and PMA2) encoding H<sup>+</sup>-ATPases. In contrast, plants have a larger number of genes for H<sup>+</sup>-ATPases. In <i>Ustilago maydis</i>, a biotrophic basidiomycete that infects corn and teosinte, the presence of two H<sup>+</sup>-ATPase-encoding genes has been described, one with high identity to the fungal enzymes (<i>pma1,</i> <i>UMAG_02851</i>), and the other similar to the plant H<sup>+</sup>-ATPases (<i>pma2,</i> <i>UMAG_01205</i>). Unlike <i>S. cerevisiae</i>, these two genes are expressed jointly in <i>U. maydis</i> sporidia. In the present work, mutants lacking one of these genes (Δ<i>pma1</i> and Δ<i>pma2</i>) were used to characterize the role of each one of these enzymes in <i>U. maydis</i> physiology and to obtain some of their kinetic parameters. To approach this goal, classical biochemical assays were performed. The absence of any of these H<sup>+</sup>-ATPases did not affect the growth or fungal basal metabolism. Membrane potential tests showed that the activity of a single H<sup>+</sup>-ATPase was enough to maintain the proton-motive force. Our results indicated that in <i>U. maydis</i>, both H<sup>+</sup>-ATPases work jointly in the generation of the electrochemical proton gradient, which is important for secondary transport of metabolites and regulation of intracellular pH. |
| first_indexed | 2024-03-09T23:22:12Z |
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| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-09T23:22:12Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
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| spelling | doaj.art-2a3f5305955440899894e3c4d5c72f5d2023-11-23T17:23:56ZengMDPI AGJournal of Fungi2309-608X2022-05-018655010.3390/jof8060550Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i>Melissa Vázquez-Carrada0Michael Feldbrügge1Dario Rafael Olicón-Hernández2Guadalupe Guerra-Sánchez3Juan Pablo Pardo4Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N Santo Tomás, Ciudad de Mexico C.P. 11340, MexicoInstitute for Microbiology, Cluster of Excellence on Plant Sciences, Heinrich Heine University Düsseldorf, 40204 Düsseldorf, GermanyDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N Santo Tomás, Ciudad de Mexico C.P. 11340, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N Santo Tomás, Ciudad de Mexico C.P. 11340, MexicoDepartamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de Mexico, Circuito Interior S/N, Ciudad Universitaria, Ciudad de Mexico C.P. 04510, MexicoPlasma membrane H<sup>+</sup>-ATPases of fungi, yeasts, and plants act as proton pumps to generate an electrochemical gradient, which is essential for secondary transport and intracellular pH maintenance. <i>Saccharomyces cerevisiae</i> has two genes (PMA1 and PMA2) encoding H<sup>+</sup>-ATPases. In contrast, plants have a larger number of genes for H<sup>+</sup>-ATPases. In <i>Ustilago maydis</i>, a biotrophic basidiomycete that infects corn and teosinte, the presence of two H<sup>+</sup>-ATPase-encoding genes has been described, one with high identity to the fungal enzymes (<i>pma1,</i> <i>UMAG_02851</i>), and the other similar to the plant H<sup>+</sup>-ATPases (<i>pma2,</i> <i>UMAG_01205</i>). Unlike <i>S. cerevisiae</i>, these two genes are expressed jointly in <i>U. maydis</i> sporidia. In the present work, mutants lacking one of these genes (Δ<i>pma1</i> and Δ<i>pma2</i>) were used to characterize the role of each one of these enzymes in <i>U. maydis</i> physiology and to obtain some of their kinetic parameters. To approach this goal, classical biochemical assays were performed. The absence of any of these H<sup>+</sup>-ATPases did not affect the growth or fungal basal metabolism. Membrane potential tests showed that the activity of a single H<sup>+</sup>-ATPase was enough to maintain the proton-motive force. Our results indicated that in <i>U. maydis</i>, both H<sup>+</sup>-ATPases work jointly in the generation of the electrochemical proton gradient, which is important for secondary transport of metabolites and regulation of intracellular pH.https://www.mdpi.com/2309-608X/8/6/550H<sup>+</sup>-ATPasesP-type ATPasesplasma membraneproton pump ATPase<i>Ustilago maydis</i> |
| spellingShingle | Melissa Vázquez-Carrada Michael Feldbrügge Dario Rafael Olicón-Hernández Guadalupe Guerra-Sánchez Juan Pablo Pardo Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i> Journal of Fungi H<sup>+</sup>-ATPases P-type ATPases plasma membrane proton pump ATPase <i>Ustilago maydis</i> |
| title | Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i> |
| title_full | Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i> |
| title_fullStr | Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i> |
| title_full_unstemmed | Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i> |
| title_short | Functional Analysis of the Plasma Membrane H<sup>+</sup>-ATPases of <i>Ustilago maydis</i> |
| title_sort | functional analysis of the plasma membrane h sup sup atpases of i ustilago maydis i |
| topic | H<sup>+</sup>-ATPases P-type ATPases plasma membrane proton pump ATPase <i>Ustilago maydis</i> |
| url | https://www.mdpi.com/2309-608X/8/6/550 |
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