title: Resistance through inhibition: ectopic expression of serine protease inhibitor offers stress tolerance via delayed senescence in yeast cell.
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.
authors.affiliation: Plant Molecular Biology Unit, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Dr. HomiBhabha Road, Pune 411 008, Maharashtra, India.
authors.forename: Rakesh S
authors.initials: RS
authors.lastname: Joshi
authors.affiliation: Plant Molecular Biology Unit, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Dr. HomiBhabha Road, Pune 411 008, Maharashtra, India.
authors.forename: Rahul S
authors.initials: RS
authors.lastname: Tanpure
authors.affiliation: Plant Molecular Biology Unit, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Dr. HomiBhabha Road, Pune 411 008, Maharashtra, India.
authors.forename: Rajan Kumar
authors.initials: RK
authors.lastname: Singh
authors.affiliation: Plant Molecular Biology Unit, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Dr. HomiBhabha Road, Pune 411 008, Maharashtra, India.
authors.forename: Vidya S
authors.initials: VS
authors.lastname: Gupta
authors.affiliation: Plant Molecular Biology Unit, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Dr. HomiBhabha Road, Pune 411 008, Maharashtra, India. Electronic address: ap.giri@ncl.res.in.
authors.forename: Ashok P
authors.initials: AP
authors.lastname: Giri
mesh_terms: D000222:Adaptation, Physiological; D002212:Capsicum / Q000737:chemistry / Q000378:metabolism; D020169:Caspases / Q000737:chemistry* / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D019216:Metals, Heavy / Q000633:toxicity; D050296:Microbial Viability; D062105:Molecular Docking Simulation; D010843:Pichia / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D054712:Salinity; D015842:Serine Proteinase Inhibitors / Q000096:biosynthesis / Q000235:genetics*; D015398:Signal Transduction; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological
chemical_list: D019216:Metals, Heavy; D029701:Saccharomyces cerevisiae Proteins; D015842:Serine Proteinase Inhibitors; D012965:Sodium Chloride; D006861:Hydrogen Peroxide; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D020169:Caspases; C456272:MCA1 protein, S cerevisiae
keywords: Metacaspase; Multiple stress tolerance; Protease inhibitor; Yeast