Characterization of putative acetate kinase in the pathogenic yeast, Cryptococcus neoformans

WCU Author/Contributor (non-WCU co-authors, if there are any, appear on document)
Beth Ann Budden (Creator)
Western Carolina University (WCU )
Web Site:
Indrani Bose

Abstract: The spherical, encapsulated basidiomycetous yeast, Cryptococcus neoformans, is an environmental opportunistic pathogen that has become a leading cause of mortality secondary to HIV/AIDS, particularly in sub-Saharan Africa (Park et al. 2009). Few chemotherapeutic agents are currently available to treat cryptococcosis, and growing concerns over resistance to some of these medications have emphasized the need for alternative treatments. By obtaining a thorough understanding of the metabolic pathways involved in the survival and pathogenicity of this organism it is hoped that pathways and/or proteins unique to the organism can be used as potential targets for chemotherapeutic agents (Casadevall and Perfect 1998). A homolog of an enzyme previously thought to exist only in bacteria, acetate kinase (AK), has been identified in certain lower eukaryotes including C. neoformans. To date, an acetate kinase homolog has not been found in higher eukaryotic organisms. To get a better understanding of the role acetate kinase plays in C. neoformans, pre-constructed ACK1 deletion strains (ack1?) were confirmed via cDNA analysis and Southern blot and were used in a variety of phenotype characterization studies. No phenotypic difference between the wild type and ack1? strains were observed. A plasmid designed for use in complementation studies was constructed but not utilized due to the absence of phenotypic variation. Several strains containing mCherry-tagged ACK1 were created and initial PCRs of the 5’ and 3’ ends suggest proper placement of the ACK1-mCherry cassette within the C. neoformans genome. Following additional placement confirmation studies, these strains can be used to identify subcellular localization of this protein and allow purification of the endogenous protein from cell lysate that can be used for enzyme kinetic studies as well as for identifying possible interacting proteins. Further exploration is needed to elucidate acetate kinase’s role in this microorganism.

Additional Information

Language: English
Date: 2013
acetate kinase, Cryptococcus neoformans
Cryptococcus neoformans -- Physiology
Cryptococcus neoformans -- Genetics

Email this document to