The Aspergillus fumigatus metacaspases CasA and CasB facilitate growth under conditions of endoplasmic reticulum stress.
We have examined the contribution of metacaspases to the growth and stress response of the opportunistic human mould pathogen, Aspergillus fumigatus, based on increasing evidence implicating the yeast metacaspase Yca1p in apoptotic-like programmed cell death. Single metacaspase-deficient mutants were constructed by targeted disruption of each of the two metacaspase genes in A. fumigatus, casA and casB, and a metacaspase-deficient mutant, DeltacasA/DeltacasB, was constructed by disrupting both genes. Stationary phase cultures of wild-type A. fumigatus were associated with the appearance of typical markers of apoptosis, including elevated proteolytic activity against caspase substrates, phosphatidylserine exposure on the outer leaflet of the membrane, and loss of viability. By contrast, phosphatidylserine exposure was not observed in stationary phase cultures of the DeltacasA/DeltacasB mutant, although caspase activity and viability was indistinguishable from wild type. The mutant retained wild-type virulence and showed no difference in sensitivity to a range of pro-apoptotic stimuli that have been reported to initiate yeast apoptosis. However, the DeltacasA/DeltacasB mutant showed a growth detriment in the presence of agents that disrupt endoplasmic reticulum homeostasis. These findings demonstrate that metacaspase activity in A. fumigatus contributes to the apoptotic-like loss of membrane phospholipid asymmetry at stationary phase, and suggest that CasA and CasB have functions that support growth under conditions of endoplasmic reticulum stress.
Richie
Daryl L
DL
Department of Pathology and Laboratory Medicine, University of Cincinnati College of Medicine, 231 Albert Sabin Way, Cincinnati, OH 45267-0529, USA.
Miley
Michael D
MD
Bhabhra
Ruchi
R
Robson
Geoffrey D
GD
Rhodes
Judith C
JC
Askew
David S
DS
D000595:Amino Acid Sequence; D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D001228:Aspergillosis; D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity; D020169:Caspases / Q000378:metabolism*; D003847:Deoxyglucose / Q000494:pharmacology; D004195:Disease Models, Animal; D004229:Dithiothreitol / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000187:drug effects*; D005260:Female; D005656:Fungal Proteins / Q000378:metabolism*; D017353:Gene Deletion; D051379:Mice; D050296:Microbial Viability; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010718:Phosphatidylserines / Q000032:analysis; D016019:Survival Analysis; D014415:Tunicamycin / Q000494:pharmacology; D014774:Virulence
D000935:Antifungal Agents; D005656:Fungal Proteins; D010718:Phosphatidylserines; D014415:Tunicamycin; D003847:Deoxyglucose; D020169:Caspases; D004229:Dithiothreitol