RNA binding protein Pub1p regulates glycerol production and stress tolerance by controlling Gpd1p activity during winemaking

Appl Microbiol Biotechnol. 2016 Jun;100(11):5017-27. doi: 10.1007/s00253-016-7340-z. Epub 2016 Feb 4.

Abstract

Glycerol is a key yeast metabolite in winemaking because it contributes to improve the organoleptic properties of wine. It is also a cellular protective molecule that enhances the tolerance of yeasts to osmotic stress and promotes longevity. Thus, its production increases by genetic manipulation, which is of biotechnological and basic interest. Glycerol is produced by diverting glycolytic glyceraldehyde-3-phosphate through the action of glycerol-3-phosphate dehydrogenase (coded by genes GPD1 and GPD2). Here, we demonstrate that RNA-binding protein Pub1p regulates glycerol production by controlling Gpd1p activity. Its deletion does not alter GPD1 mRNA levels, but protein levels and enzymatic activity increase, which explains the higher intracellular glycerol concentration and greater tolerance to osmotic stress of the pub1∆ mutant. PUB1 deletion also enhances the activity of nicotinamidase, a longevity-promoting enzyme. Both enzymatic activities are partially located in peroxisomes, and we detected peroxisome formation during wine fermentation. The role of Pub1p in life span control depends on nutrient conditions and is related with the TOR pathway, and a major connection between RNA metabolism and the nutrient signaling response is established.

Keywords: Glycerol; Gpd1p; Peroxisomes; Pub1p; Saccharomyces cerevisiae; TOR1/Sch9 nutrient signaling pathway; Winemaking.

MeSH terms

  • Fermentation
  • Food Handling
  • Fruit and Vegetable Juices / microbiology
  • Gene Expression Regulation, Fungal
  • Glycerol / metabolism*
  • Glycerol-3-Phosphate Dehydrogenase (NAD+) / genetics
  • Glycerol-3-Phosphate Dehydrogenase (NAD+) / metabolism*
  • Glycerolphosphate Dehydrogenase / genetics
  • Glycerolphosphate Dehydrogenase / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Membrane Transport Proteins / genetics
  • Membrane Transport Proteins / metabolism
  • Osmotic Pressure
  • Phosphatidylinositol 3-Kinases / genetics
  • Phosphatidylinositol 3-Kinases / metabolism
  • Poly(A)-Binding Proteins / genetics
  • Poly(A)-Binding Proteins / metabolism*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Stress, Physiological
  • Vitis / microbiology
  • Wine / analysis*
  • Wine / microbiology

Substances

  • FPS1 protein, S cerevisiae
  • Membrane Proteins
  • Membrane Transport Proteins
  • PUB1 protein, S cerevisiae
  • Poly(A)-Binding Proteins
  • RNA, Messenger
  • STL1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Glycerolphosphate Dehydrogenase
  • GPD1 protein, S cerevisiae
  • GPD2 protein, S cerevisiae
  • Glycerol-3-Phosphate Dehydrogenase (NAD+)
  • TOR1 protein, S cerevisiae
  • Protein Serine-Threonine Kinases
  • SCH9 protein, S cerevisiae
  • Glycerol