title: Leishmania major metacaspase can replace yeast metacaspase in programmed cell death and has arginine-specific cysteine peptidase activity.
abstract: The human protozoan parasite Leishmania major has been shown to exhibit several morphological and biochemical features characteristic of a cell death program when differentiating into infectious stages and under a variety of stress conditions. Although some caspase-like peptidase activity has been reported in dying parasites, no caspase gene is present in the genome. However, a single metacaspase gene is present in L. major whose encoded protein harbors the predicted secondary structure and the catalytic dyad histidine/cysteine described for caspases and other metacaspases identified in plants and yeast. The Saccharomyces cerevisiae metacaspase YCA1 has been implicated in the death of aging cells, cells defective in some biological functions, and cells exposed to different environmental stresses. In this study, we describe the functional heterologous complementation of a S. cerevisiae yca1 null mutant with the L. major metacaspase (LmjMCA) in cell death induced by oxidative stress. We show that LmjMCA is involved in yeast cell death, similar to YCA1, and that this function depends on its catalytic activity. LmjMCA was found to be auto-processed as occurs for caspases, however LmjMCA did not exhibit any activity with caspase substrates. In contrast and similarly to Arabidopsis thaliana metacaspases, LmjMCA was active towards substrates with arginine in the P1 position, with the activity being abolished following H147A and C202A catalytic site mutations. These results suggest that metacaspases are members of a family of peptidases with a role in cell death conserved in evolution notwithstanding possible differences in their catalytic activity.
authors.affiliation: Department of Biochemistry, University of Lausanne, 155 Chemin des Boveresses, CH-1066 Epalinges, Switzerland.
authors.forename: Iveth J
authors.initials: IJ
authors.lastname: González
authors.forename: Chantal
authors.initials: C
authors.lastname: Desponds
authors.forename: Cédric
authors.initials: C
authors.lastname: Schaff
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Mottram
authors.forename: Nicolas
authors.initials: N
authors.lastname: Fasel
mesh_terms: D000818:Animals; D001120:Arginine / Q000737:chemistry*; D020169:Caspases / Q000737:chemistry*; D016923:Cell Death; D003545:Cysteine / Q000737:chemistry*; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D006801:Humans; D018320:Leishmania major / Q000235:genetics / Q000378:metabolism*; D015003:Yeasts / Q000378:metabolism*
chemical_list: D001120:Arginine; D020169:Caspases; D003546:Cysteine Endopeptidases; D003545:Cysteine