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• W1897968124 abstract "Cryptococcus neoformans is a basidiomycetous yeast with clinical importancedue to its role as an opportunistic pathogen in immunocompromised individuals. Uponexposure to C. neoformans patients may develop a disease known as cryptococcosis, acondition associated with pulmonary infections that may result in lethalmeningoencephalitis. This disease is often accompanied by aneuploidies of single ormultiple chromosomes in a subpopulation of the infecting cells, leading to the idea thatmissegregation of chromosomes during infection may be a way for this organism to dealwith stresses and acquiring antifungal resistance. Because this species is rapidlydeveloping resistance to many of the prevalent antifungal treatments, a need exists formore potent therapies with fewer side effects. The molecular biology concerning cellularreproduction and division in C. neoformans is poorly understood for such an importantfungal pathogen. The research performed so far has produced information about suchstructures as the centromeric DNA of their chromosomes and the kinetochore proteinsuperstructure that links the spindle microtubules to the centromeres. Understanding howchromosome segregation occurs in C. neoformans may bring about better therapies: if itcan be shown that homologous proteins perform the same actions in Cryptococcus as they do in other organisms then these proteins may potentially become targets for futurestudies to determine the mechanism linking the presence of antifungal chemicals toaneuploidy.The kinetochore protein complex is essential for segregation of chromosomes.Previous experiments have described a protein essential to the structure and function ofthe yeast kinetochore. The necessity of this protein for kinetochore function was firstdetermined using an Schizosaccharomyces pombe strain carrying a temperature-sensitivemutation in the Mis12 gene that causes the protein to malfunction above a certaintemperature. It was found that when grown above the restrictive temperature cells wereunable to complete mitotic segregation, with the dividing nucleus stuck in the neckbetween budding cells. A later study described the budding yeast homolog MTW1(Mis12-like protein) in Saccharomyces cerevisiae using a similar temperature-sensitivemutant and yielded similar results. This protein is part of a subcomplex of kinetochoreproteins called the MIND complex. One goal of this project is to characterize the MTW1homolog in C. neoformans in order to determine whether it maintains the functionalimportance to kinetochore structure exhibited by the previously described homologs.This has been addressed through the construction of similar temperature-sensitivemutants and by regulating the gene expression through regulatable CTR4 promoter.These strains were created through the use of overlap PCR and biolistic transformation.Molecular and phenotypic characterizations have been performed on these strains andinclude growth and morphological analyses. Results have confirmed the presence of thedesired mutations and have shown some inhibitory effect on growth under restrictive conditions. RNA interference was performed in an effort to demonstrate the necessity of various kinetochore proteins. Initial results indicate that MTW1, along with the outer kinetochore gene DAD2, are not essential whereas several other MIND complex genes (NNF1, DSN1) and inner (CSE4, MIF2) and outer (DAD1) kinetochore genes were shownto be necessary for viability of the yeast cells. These results are in accordance with thoseobtained through the use of strains with conditional mutations in MTW1. These resultsimply that while CnMTW1 probably acts in the same capacity as that of its ascomycetoushomologs and is necessary for optimal kinetochore function, it is not essential forchromosome segregation." @default.
• W1897968124 created "2016-06-24" @default.
• W1897968124 creator A5026802113 @default.
• W1897968124 date "2013-01-01" @default.
• W1897968124 modified "2023-09-27" @default.
• W1897968124 title "Characterizing the role of MTW1 in the kinetochore of the pathogenic yeast Cryptococcus neoformans" @default.
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