Interaction between the plant ApDef1 defensin and Saccharomyces cerevisiae results in yeast death through a cell cycle- and caspase-dependent process occurring via uncontrolled oxidative stress

被引：42

作者：

Soares, Julia Ribeiro ^{[1
]}

Tenorio de Melo, Edesio Jose ^{[2
]}

da Cunha, Maura ^{[2
]}

Sales Fernandes, Katia Valevski ^{[3
]}

Taveira, Gabriel Bonan ^{[1
]}

Pereira, Lidia da Silva ^{[4
]}

Pimenta, Samy ^{[4
]}

Trindade, Fernanda Gomes ^{[2
]}

Regente, Mariana ^{[5
]}

Pinedo, Marcela ^{[5
]}

de la Canal, Laura ^{[5
]}

Gomes, Valdirene Moreira ^{[1
]}

Carvalho, Andre de Oliveira ^{[1
]}

机构：

[1] Univ Estadual Norte Fluminense, Ctr Biociencias & Biotecnol, Lab Fisiol & Bioquim Microrganismos, Parque Calif, BR-28013602 Campos Dos Goytacazes, RJ, Brazil

[2] Univ Estadual Norte Fluminense, Ctr Biociencias & Biotecnol, Lab Biol Celular & Tecidual, Campos Dos Goytacazes, RJ, Brazil

[3] Univ Estadual Norte Fluminense, Lab Quim & Funcao Prot & Peptideos, Campos Dos Goytacazes, RJ, Brazil

[4] Univ Estadual Norte Fluminense, Ctr Ciencias & Tecnol Agr, Lab Melhoramento Genet Vegetal, Campos Dos Goytacazes, RJ, Brazil

[5] Univ Nacl Mar del Plata, CONICET, Inst Invest Biol, Mar Del Plata, Buenos Aires, Argentina

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 2017年 / 1861卷 / 01期

关键词：

Antimicrobial peptides; Microbicidal; Membrane permeabilization; ROS induction; Cell cycle; Caspase; ANTIMICROBIAL PEPTIDES; CANDIDA-ALBICANS; RAPHANUS-SATIVUS; SPERMIDINE TRANSPORT; ESCHERICHIA-COLI; APOPTOSIS; MECHANISMS; MEMBRANE; IDENTIFICATION; INDUCTION;

D O I：

10.1016/j.bbagen.2016.09.005

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Background: Plant defensins were discovered at beginning of the 90s'; however, their precise mechanism of action is still unknown. Herein, we studied ApDef(1)-Saccharomyces cerevisiae interaction. Methods: ApDef(1)-S. cerevisiae interaction was studied by determining the MIC, viability and death kinetic assays. Viability assay was repeated with hydroxyurea synchronized-yeast and pretreated with CCCP. Plasma membrane permeabilization, ROS induction, chromatin condensation, and caspase activation analyses were assessed through Sytox green, DAB, DAPI and FITC-VAD-FMK, respectively. Viability assay was done in presence of ascorbic acid and Z-VAD-FMK. Ultrastructural analysis was done by electron microscopy. Results: ApDef(1) caused S. cerevisiae cell death and MIC was 7.8 mu M. Whole cell population died after 18 h of ApDef(1) interaction. After 3 h, 98.76% of synchronized cell population died. Pretreatment with CCCP protected yeast from ApDef(1) induced death. ApDef(1)-S. cerevisiae interaction resulted in membrane permeabilization, H2O2 increased production, chromatin condensation and caspase activation. Ascorbic acid prevented yeast cell death and membrane permeabilization. Z-VAD-FMK prevented yeast cell death. Conclusions: ApDef(1)-S. cerevisiae interaction caused cell death through cell cycle dependentprocess which requires preserved membrane potential. After interaction, yeast went through uncontrolled ROS production and accumulation, which led to plasma membrane permeabilization, chromatin condensation and, ultimately, cell death by activation of caspase-dependent apoptosis via. General significance: We show novel requirements for the interaction between plant defensin and fungi cells, i.e. cell cycle phase and membrane potential, and we indicate that membrane permeabilization is probably caused by ROS and therefore, it would be an indirect event of the ApDef(1)-S. cerevisiae interaction. (C) 2016 Elsevier B.V. All rights reserved.

引用

页码：3429 / 3443

页数：15

共 92 条

[1] The antifungal activity of RsAFP2, a plant defensin from Raphanus sativus, involves the induction of reactive oxygen species in Candida albicans [J].