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)
- Institution
- Western Carolina University (WCU )
- Web Site: http://library.wcu.edu/
- Advisor
- 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.
Characterization of putative acetate kinase in the pathogenic yeast, Cryptococcus neoformans
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Created on 4/1/2013
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Additional Information
- Publication
- Thesis
- Language: English
- Date: 2013
- Keywords
- acetate kinase, Cryptococcus neoformans
- Subjects
- Cryptococcus neoformans -- Physiology
- Cryptococcus neoformans -- Genetics