caa a22 4500 605499632 CHVBK 20210128100552.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6708-9 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6708-9 PEP3 overexpression shortens lag phase but does not alter growth rate in Saccharomyces cerevisiae exposed to acetic acid stress [Elektronische Daten] [Jun Ding, Garrett Holzwarth, C. Bradford, Ben Cooley, Allen Yoshinaga, Jana Patton-Vogt, Hagai Abeliovich, Michael Penner, Alan Bakalinsky] In fungi, two recognized mechanisms contribute to pH homeostasis: the plasma membrane proton-pumping ATPase that exports excess protons and the vacuolar proton-pumping ATPase (V-ATPase) that mediates vacuolar proton uptake. Here, we report that overexpression of PEP3 which encodes a component of the HOPS and CORVET complexes involved in vacuolar biogenesis, shortened lag phase in Saccharomyces cerevisiae exposed to acetic acid stress. By confocal microscopy, PEP3-overexpressing cells stained with the vacuolar membrane-specific dye, FM4-64 had more fragmented vacuoles than the wild-type control. The stained overexpression mutant was also found to exhibit about 3.6-fold more FM4-64 fluorescence than the wild-type control as determined by flow cytometry. While the vacuolar pH of the wild-type strain grown in the presence of 80mM acetic acid was significantly higher than in the absence of added acid, no significant difference was observed in vacuolar pH of the overexpression strain grown either in the presence or absence of 80mM acetic acid. Based on an indirect growth assay, the PEP3-overexpression strain exhibited higher V-ATPase activity. We hypothesize that PEP3 overexpression provides protection from acid stress by increasing vacuolar surface area and V-ATPase activity and, hence, proton-sequestering capacity. Springer-Verlag Berlin Heidelberg, 2015 Saccharomyces cerevisiae nationallicence Yeast nationallicence Acetic acid nationallicence PEP3 nationallicence V-ATPase nationallicence HOPS nationallicence CORVET nationallicence Vacuole nationallicence STM1 nationallicence PEP5 nationallicence Ding Jun Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA aut Holzwarth Garrett Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA aut Bradford C. Department of Environmental & Molecular Toxicology, Oregon State University, Corvallis, OR, USA aut Cooley Ben Biological Sciences, Duquesne University, Pittsburgh, PA, USA aut Yoshinaga Allen Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA aut Patton-Vogt Jana Biological Sciences, Duquesne University, Pittsburgh, PA, USA aut Abeliovich Hagai Department of Biochemistry and Food Science, Hebrew University, Rehovot, Israel aut Penner Michael Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA aut Bakalinsky Alan Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8667-8680 0175-7598 99:20<8667 2015 99 253 https://doi.org/10.1007/s00253-015-6708-9 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/s00253-015-6708-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ding Jun Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Holzwarth Garrett Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Bradford C. Department of Environmental & Molecular Toxicology, Oregon State University, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Cooley Ben Biological Sciences, Duquesne University, Pittsburgh, PA, USA aut NATIONALLICENCE 700 1- Yoshinaga Allen Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Patton-Vogt Jana Biological Sciences, Duquesne University, Pittsburgh, PA, USA aut NATIONALLICENCE 700 1- Abeliovich Hagai Department of Biochemistry and Food Science, Hebrew University, Rehovot, Israel aut NATIONALLICENCE 700 1- Penner Michael Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Bakalinsky Alan Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8667-8680 0175-7598 99:20<8667 2015 99 253