Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Role of VTC4 in Stress Response and Regulation of Inorganic Polyphosphate Levels in Yeast

Version 1 : Received: 21 October 2020 / Approved: 23 October 2020 / Online: 23 October 2020 (10:42:43 CEST)

How to cite: Tomashevsky, A.; Kulakovskaya, E.; Trilisenko, L.; Kulakovskaya, T.; Fedorov, A.; Eldarov, M. Role of VTC4 in Stress Response and Regulation of Inorganic Polyphosphate Levels in Yeast. Preprints 2020, 2020100484 (doi: 10.20944/preprints202010.0484.v1). Tomashevsky, A.; Kulakovskaya, E.; Trilisenko, L.; Kulakovskaya, T.; Fedorov, A.; Eldarov, M. Role of VTC4 in Stress Response and Regulation of Inorganic Polyphosphate Levels in Yeast. Preprints 2020, 2020100484 (doi: 10.20944/preprints202010.0484.v1).

Abstract

Inorganic polyphosphate (polyP) is an important factor of stress tolerance in microbial cells. In yeast, the major enzyme of polyP biosynthesis is Vtc4, a subunit of the vacuole transporter chaperone (VTC) complex. In this study, we demonstrated that Vtc4 knockout in Saccharomyces cerevisiae not only decreased polyP content but also caused shifts in the composition of the intracellular polyP pool and changed the stress tolerance profile. In the mutant S. cerevisiae, the level of short-chain acid-soluble polyPs was decreased nearly 10-fold, whereas that of longer acid-insoluble polyPs was decreased only 2-fold, suggesting the existence of other enzymes compensating the production of long-chain polyPs. The Δvtc4 mutant showed inhibition of Mg2+-dependent phosphate uptake and decreased resistance to alkaline stress but increased tolerance to oxidation and heavy metal ions, especially Mn2+. Quantitative PCR revealed the upregulation of the DDR2 gene implicated in multiple stress responses and downregulation of PHO84 encoding a phosphate and Mn2+ transporter, which could account for the effects on phosphate uptake and Mn2+-related stress response in the Δvtc4 mutant. Our study indicates that short-chain polyPs, plays an important role in the regulation of stress response in yeast.

Subject Areas

inorganic polyphosphate; VTC4; knockout mutant; oxidative stress; gene expression; yeast

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