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dc.contributor.advisorMichael R. Green
dc.contributor.authorApone, Lynne Marie
dc.date2022-08-11T08:08:39.000
dc.date.accessioned2022-08-23T16:03:28Z
dc.date.available2022-08-23T16:03:28Z
dc.date.issued1998-01-14
dc.date.submitted2006-09-13
dc.identifier.doi10.13028/31jk-tj98
dc.identifier.urihttp://hdl.handle.net/20.500.14038/31484
dc.description<p>Some images did not scan well. Please consult original document.</p>
dc.description.abstractTranscription by RNA polymerase II is a highly regulated process requiring a number of general and promoter specific transcription factors. Although many of the factors involved in the transcription reaction are known, exactly how they function to stimulate or repress transcription is not well understood. Central to understanding gene regulation is understanding the mechanism by which promoter specific transcription activators (activators) stimulate transcription. A group of factors called coactivators have been shown to be required for activator function in vitro. The best characterized coactivators to date are members of the TFIID complex. TFIID is a multisubunit complex composed of the TATA box binding protein (TBP) and 8-12 TBP associated factors (TAFIIs). Results from numerous in vitro experiments indicate that TAFIIs function by binding to activators and forming a bridge between the activator and the basal transcription machinery. In order to gain insight into the mechanism by which activators stimulate transcription, we chose to analyze the in vivo function of TAFIIs, their proposed targets. Results from the genetic disruption of a number of TAFIIs in the yeast Saccharomyces cerevisiae showed that most are encoded by essential genes. In order to study their function, temperature-sensitive and conditional alleles were constructed. Cells depleted of individual TAFIIs by either of these two methods displayed no defect in global transcription activation. Inactivation of yTAFII17, however, resulted in a promoter specific defect. In addition, inactivation of yTAFII145, yTAFII90, or TSM1, resulted in an inability of cells to progress through the cell-cycle. In an attempt to identify genes whose expression required yTAFII90, we performed subtractive hybridization on strains containing wild-type and temperature-sensitive alleles. Although this technique successfully identified genes differentially expressed in the two strains, it failed to identify genes whose expression required yTAFII90. These results indicate that TAFIIs are not the obligatory targets of activators, and that other factors must provide this role in vivo. Furthermore, that many of TAFIIs are required for cell-cycle progression.
dc.language.isoen_US
dc.rightsCopyright is held by the author, with all rights reserved.
dc.subjectTrans-Activation (Genetics)
dc.subjectTranscription Factors
dc.subjectGene Expression Regulation
dc.subjectRNA Polymerase II
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectFungi
dc.subjectGenetic Phenomena
dc.subjectGenetics
dc.subjectNucleic Acids, Nucleotides, and Nucleosides
dc.titleAnalysis of TAF II Function in the Yeast Saccharomyces Cerevisiae
dc.typeDoctoral Dissertation
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1142&amp;context=gsbs_diss&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/gsbs_diss/183
dc.legacy.embargo2017-04-24T00:00:00-07:00
dc.identifier.contextkey205183
refterms.dateFOA2022-08-27T04:44:00Z
html.description.abstract<p>Transcription by RNA polymerase II is a highly regulated process requiring a number of general and promoter specific transcription factors. Although many of the factors involved in the transcription reaction are known, exactly how they function to stimulate or repress transcription is not well understood. Central to understanding gene regulation is understanding the mechanism by which promoter specific transcription activators (activators) stimulate transcription.</p> <p>A group of factors called coactivators have been shown to be required for activator function <em>in vitro</em>. The best characterized coactivators to date are members of the TFIID complex. TFIID is a multisubunit complex composed of the TATA box binding protein (TBP) and 8-12 TBP associated factors (TAF<sub>II</sub>s). Results from numerous <em>in vitro</em> experiments indicate that TAF<sub>II</sub>s function by binding to activators and forming a bridge between the activator and the basal transcription machinery. In order to gain insight into the mechanism by which activators stimulate transcription, we chose to analyze the <em>in vivo</em> function of TAF<sub>II</sub>s, their proposed targets.</p> <p>Results from the genetic disruption of a number of TAF<sub>II</sub>s in the yeast <em>Saccharomyces cerevisiae</em> showed that most are encoded by essential genes. In order to study their function, temperature-sensitive and conditional alleles were constructed. Cells depleted of individual TAF<sub>II</sub>s by either of these two methods displayed no defect in global transcription activation. Inactivation of yTAF<sub>II</sub>17, however, resulted in a promoter specific defect. In addition, inactivation of yTAF<sub>II</sub>145, yTAF<sub>II</sub>90, or TSM1, resulted in an inability of cells to progress through the cell-cycle.</p> <p>In an attempt to identify genes whose expression required yTAF<sub>II</sub>90, we performed subtractive hybridization on strains containing wild-type and temperature-sensitive alleles. Although this technique successfully identified genes differentially expressed in the two strains, it failed to identify genes whose expression required yTAF<sub>II</sub>90.</p> <p>These results indicate that TAF<sub>II</sub>s are not the obligatory targets of activators, and that other factors must provide this role <em>in vivo</em>. Furthermore, that many of TAF<sub>II</sub>s are required for cell-cycle progression.</p>
dc.identifier.submissionpathgsbs_diss/183
dc.contributor.departmentMolecular, Cell and Cancer Biology
dc.description.thesisprogramBiochemistry and Molecular Pharmacology


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