Resistance through inhibition: ectopic expression of serine protease inhibitor offers stress tolerance via delayed senescence in yeast cell

Biochem Biophys Res Commun. 2014 Sep 26;452(3):361-8. doi: 10.1016/j.bbrc.2014.08.075. Epub 2014 Aug 23.

Abstract

Protease inhibitors have been known to confer multiple stress tolerance in transgenic plants. We have assessed growth of yeast (Pichia pastoris GS115) strains expressing inhibitory repeat domains (PpIRD(+)) of previously characterized Capsicum annuum protease inhibitors under high salt, heavy metal and oxidative stress. PpIRD(+) strains exhibited multiple stress tolerance and showed differential molecular responses at transcriptional and translational level on exposure to stress inducing agents like heavy metal, high salt and H2O2. PpIRD(+) strains display significant reduction in metacaspase (Yca1) activity, the key enzyme in apoptosis, indicates the possibility of cross reactivity of IRDs (serine protease inhibitor) with cysteine proteases. PpIRD(+) and Saccharomyces cerevisiae knockout with Yca1 (ΔYca1) strain showed similar growth characteristics under stress, which indicated the delayed senescence due to cellular metacaspase inhibition. Molecular docking study showed a close proximity of IRDs reactive site and the active site of metacaspase in the complex that signified their strong interactions. Maintenance of GAPDH activity, primary target of metacaspase, in PpIRD(+) strain evidenced the inhibition of metacaspase activity and survival of these cells under stress. This report demonstrates a potential molecular mechanism of protease inhibitor-based multiple stress tolerance in yeast strains.

Keywords: Metacaspase; Multiple stress tolerance; Protease inhibitor; Yeast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptation, Physiological
  • Capsicum / chemistry
  • Capsicum / metabolism
  • Caspases / chemistry*
  • Caspases / deficiency
  • Caspases / genetics
  • Caspases / metabolism
  • Gene Expression Regulation, Fungal*
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / genetics
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / metabolism
  • Hydrogen Peroxide / pharmacology
  • Metals, Heavy / toxicity
  • Microbial Viability
  • Molecular Docking Simulation
  • Pichia / drug effects
  • Pichia / genetics*
  • Pichia / metabolism
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Salinity
  • Serine Proteinase Inhibitors / biosynthesis
  • Serine Proteinase Inhibitors / genetics*
  • Signal Transduction
  • Sodium Chloride / pharmacology
  • Stress, Physiological

Substances

  • Metals, Heavy
  • Saccharomyces cerevisiae Proteins
  • Serine Proteinase Inhibitors
  • Sodium Chloride
  • Hydrogen Peroxide
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)
  • Caspases
  • MCA1 protein, S cerevisiae