Characterization of FIG2 in Saccharomyces cerevisiae.
Item
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Title
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Characterization of FIG2 in Saccharomyces cerevisiae.
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Identifier
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AAI9997099
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identifier
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9997099
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Creator
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Jue, Chong.
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Contributor
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Adviser: Peter Lipke
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Date
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2001
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Molecular
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Abstract
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The complete genome sequence of the yeast Saccharomyces cerevisiae has been available since 1996. One of the most immediate, albeit difficult, consequences of this is the ability to investigate the functions of genes that have not been previously characterized.;One such previously uncharacterized open reading frame, YCR089W /FIG2, has some of the structural characteristics of a cell wall mannoprotein. Construction of a fusion protein consisting of this open reading frame and jellyfish green-fluorescence-protein indicated localization to the cell wall.;Mutant strains were constructed in which this gene was either deleted or overexpressed. These strains were then characterized in terms of cell wall structure and function. FIG2 was found to be constitutively expressed in both a and alpha haploid mating-type cells.;With respect to the structure of the cell wall, it was found that loss of FIG2 resulted in aberrant cell morphology in cells of either mating type grown to stationary phase. When cells of either mating type were induced with the appropriate mating pheromone from the opposite type, FIG2 expression was increased. This induction of FIG2 expression was greater in alpha cells than in a cells.;With respect to functionality, the sexual agglutinability of fig2Delta alpha mating-type cells was significantly higher than the corresponding wild type strain. Conversely, the overexpression of FIG2 in alpha cells resulted in decreased agglutinability. However, when FIG2 was either deleted or overexpressed in a cells, no changes in agglutinability were observed. The time course of the pheromone induction of a cells was not altered by deletion of FIG2.;The molecular basis of the inhibition of alpha cell agglutination by FIG2 was partially characterized. Increasing the gene dosage of FIG2 did not affect the level of transcription of alpha-agglutinin. It was therefore hypothesized that the decrease in agglutinability of alpha cells overexpressing, FIG2 could be the result of specific binding of Fig2p to alpha-agglutinin. Experiments measuring the binding of soluble, labeled alpha-agglutinin, to whole alpha cells, both wild type and FIG2 delete, did not indicate any specific binding.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
