title: NP24 induces apoptosis dependent on caspase-like activity in Saccharomyces cerevisiae.
abstract: Tomato NP24 is a homolog of osmotin, a PR-5 protein from tobacco that can initiate apoptosis in yeast via PHO36 in the plasma membrane. We cloned and sequenced NP24 from tomato cv. Momotaro. Based on phylogenetic analysis, NP24 from Momotaro belonged to the Solanaceae clade. The amino acid sequence was identical to that of cv. Ailsa Craig including signal peptide, but the residues predicted to interact with the adiponectin receptor, ADIPOR, were slightly different from osmotin. Recombinant NP24 (rNP24) was expressed in a reductase-deficient mutant of Escherichia coli as host cell, and purified from cell extract by affinity chromatography. Purified rNP24 significantly inhibited growth of Saccharomyces cerevisiae wild-type spheroplasts. In contrast, growth of PHO36 deletion mutant (ΔIzh2) spheroplasts was not inhibited. Moreover, rNP24 induced significant activity of reactive oxygen species, caspase-like activity, and also nuclear fragmentation in wild-type spheroplast cells. These results demonstrated that rNP24 from Momotaro greatly influenced cell viability due to triggering apoptosis through PHO36. Notably, apoptosis induced by NP24 was caspase-like protease dependent.
authors.affiliation: Applied Microbiology and Biotechnology Laboratory, Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Higuchi
authors.affiliation: Food Biotechnology Laboratory, National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Applied Microbiology and Biotechnology Laboratory, Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kato
authors.affiliation: Applied Microbiology and Biotechnology Laboratory, Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ogihara
authors.affiliation: Applied Microbiology and Biotechnology Laboratory, Department of Chemistry and Life Science, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan. Electronic address: kasumi@brs.nihon-u.ac.jp.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Kasumi
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D018551:Solanum lycopersicum / Q000737:chemistry*; D008565:Membrane Proteins / Q000172:deficiency / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D021382:Protein Sorting Signals; D017382:Reactive Oxygen Species / Q000378:metabolism; D054419:Receptors, Adiponectin / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013104:Spheroplasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D014026:Nicotiana / Q000737:chemistry
chemical_list: C497767:IZH2 protein, S cerevisiae; D008565:Membrane Proteins; D010940:Plant Proteins; D021382:Protein Sorting Signals; D017382:Reactive Oxygen Species; D054419:Receptors, Adiponectin; D029701:Saccharomyces cerevisiae Proteins; C073139:osmotin protein, Nicotiana tabacum; D020169:Caspases
keywords: Adiponectin; Apoptosis; Caspase; NP24; Osmotin; PHO36