Open AccessThe Mcf1 toxin induces apoptosis via the mitochondrial pathway and apoptosis is attenuated by mutation of the BH3‐like domain
Author(s) -
Dowling Andrea J.,
Waterfield Nicholas R.,
Hares Michelle C.,
Le Goff Gaëlle,
Streuli Charles H.,
FfrenchConstant Richard H.
Publication year - 2007
Publication title -
cellular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.542
H-Index - 138
eISSN - 1462-5822
pISSN - 1462-5814
DOI - 10.1111/j.1462-5822.2007.00974.x
Subject(s) - biology , apoptosis , microbiology and biotechnology , mitochondrion , dna laddering , death domain , cytochrome c , effector , intrinsic apoptosis , pyrin domain , mitochondrial apoptosis induced channel , inhibitor of apoptosis domain , caspase , programmed cell death , genetics , dna fragmentation , inflammasome , receptor
Summary Photorhabdus are Gram‐negative, nematode‐vectored bacteria that produce toxins to kill their insect hosts. The expression of one of these, Makes caterpillars floppy 1 (Mcf1), is sufficient to allow Escherichia coli to survive within, and kill, caterpillars which are otherwise able to clear E. coli infection. Mcf1 treated caterpillars show rapid loss of body turgor (the ‘floppy’ phenotype) and death is associated with massive apoptosis of both the midgut epithelium and insect phagocytes. Mammalian tissue culture cells treated with Mcf1 also display key features of apoptosis including zeiosis, apoptotic nuclear morphology, DNA laddering, activation of the effector caspase‐3 and PARP cleavage. As Mcf1 carries a single BH3‐like domain, here we investigate the hypothesis that this toxin promotes apoptosis via the mitochondrial pathway by mimicking a BH3 domain‐only protein. Consistent with this hypothesis, a double mutant within the BH3‐like domain causes a dramatic decline in apoptosis. Mcf1 also alters mitochondrial membrane potential and triggers the release of cytochrome c . Cells overexpressing Bcl‐x L , an anti‐apoptotic Bcl‐2 family member, are resistant to Mcf1‐mediated apoptosis, as are cells deficient in Bax. In addition, translocation of Bax to the mitochondrion is observed in response to Mcf1 treatment. Together, these results show that Mcf1 mediates apoptosis via the mitochondrial pathway, and are consistent with the hypothesis that the BH3‐like domain in Mcf1 is a functional requirement for the pro‐apoptotic activity of Mcf1.