Metacaspase gene function in the mushroom fungus Schizophyllum commune

The overall goal of this project was to investigate the biological role of a putative metacaspase gene present in the mushroom fungus Schizophyllum commune. For this study, we have utilized a strain of S. commune that is unable to integrate DNA via the non-homologous end joining pathway. This forces transforming DNA to integrate homologously, as is required for the purposes of gene knockout. The gene Scp1 encodes a likely member of the metacaspase protein family, which are suspected to have activity similar to caspases, the latter crucial to programmed cell death. A knockout construct containing a non-functional version of Scp1 was previously generated in our laboratory. This DNA was then transformed into Schizophyllum commune in an attempt to knockout the native Scp1 gene. At present a likely knockout (null) strain has been identified, and analysis by polymerase chain reaction has supported its status as a true knockout. A homozygous null mutant of Scp1 will then be generated, and will be compared to a wild-type strain for any alterations in colony growth and/or mushroom development. The role(s) of other members of the metacaspase family will eventually be examined by a similar approach.