Advertisement

Current Genetics

, Volume 36, Issue 1–2, pp 21–28 | Cite as

Reconstitution of caspase-mediated cell-death signalling in Schizosaccharomyces pombe

  • Stephan Ryser
  • Elisabeth Vial
  • Edith Magnenat
  • Werner Schlegel
  • Kinsey Maundrell
ORIGINAL PAPER

Abstract

Two pro-apoptotic proteases, caspase-1 and caspase-3, have been expressed as full-length proteins in the fission yeast Schizosaccharomyces pombe. Both proteins autoprocess to generate the corresponding active enzyme and both are lethal to the yeast cell. Lethality is due to catalytic activity since the expression of the inactive mutant forms of both caspases does not result in an obvious phenotype. Caspase-expressing yeast can be rescued by co-expression of the baculovirus protein p35, a known inhibitor of the caspase family. Co-expression of Bcl-2, another anti-apoptotic protein, does not prevent the cell death induced by either caspase. However, Bcl-2 is itself cleaved by both caspase-1 and caspase-3 at two adjacent recognition sites, YEWD31′A and DAGD34′V respectively, immediately downstream from the N-terminal BH4 domain, a region of Bcl-2 which is essential for its anti-apoptotic activity; similar cleavage of Bcl-2 by caspases has been demonstrated in mammalian cells. Hence, key elements of the apoptotic pathway can be reliably reconstituted in fission yeast, opening the way to exploit yeast in order to study the control of apoptosis. Furthermore, the activity of caspase-3, although not caspase-1, can be demonstrated in vitro using chromogenic substrates. This offers the possibility of using caspase-producing strains of yeast to screen for chemical inhibitors either in vivo or in vitro.

Key words Fission yeast Caspases Bcl-2 Apoptosis 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Stephan Ryser
    • 1
  • Elisabeth Vial
    • 1
  • Edith Magnenat
    • 1
  • Werner Schlegel
    • 2
  • Kinsey Maundrell
    • 1
  1. 1.Ares-Serono, 14 ch. des Aulx, 1228 Plan-Les-Ouates, CH-1211 Geneva, SwitzerlandCH
  2. 2.Fondation pour Recherches Médicales, 64 av. de la Roseraie, CH-1211 Geneva 4, Switzerland e-mail: ryser0@cmu.unige.ch Tel.: +41-22-3823815 Fax: +41-22-3475979CH

Personalised recommendations