Transcriptional regulation of interleukin-2 gene expression through NF-kB and RW-BF.
Item
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Title
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Transcriptional regulation of interleukin-2 gene expression through NF-kB and RW-BF.
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Identifier
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AAI9325107
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identifier
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9325107
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Creator
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Hoyos, Beatrice.
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Contributor
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Adviser: Miriam Siekevitz
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Date
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1993
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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, Microbiology | Biology, Molecular | Health Sciences, Immunology
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Abstract
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Interleukin-2 (IL-2) is a T cell growth factor required for the clonal expansion of activated T cells. This lymphokine is regulated at the level of transcription, and several cis-elements and transacting factors that regulate its level of expression have been identified. This thesis work has concentrated on the identification and characterization of DNA binding proteins interacting with the sequences {dollar}-{dollar}206 to {dollar}-{dollar}195 and {dollar}-{dollar}197 to {dollar}-{dollar}183 upstream of the transcription initiation site in the IL-2 promoter.;Inspection of the IL-2 promoter led to the recognition of the sequence {dollar}-{dollar}206 to {dollar}-{dollar}195 as an element that shares 7 out of 10 bases with the NF-kB binding motif. Mutagenesis of this sequence resulted in 30- to 75% down regulation of the promoter function. The factor binding this motif was identified as Nuclear Factor-kappa B (NF-kB) based on several criteria.;Mutagenesis of the sequence (termed RW) situated immediately downstream of NF-kB at position {dollar}-{dollar}197 to {dollar}-{dollar}183 was conducted to determine its contribution to the regulation of IL-2 gene expression. The results showed a 25- to 50% decrease in promoter function. The binding of the complex RW-binding factor (RW-BF) was characterized and differentiated from NF-kB binding.;To identify the protein binding to this consensus sequence a HUT78 cDNA expression library was screened using as probe the sequence {dollar}-{dollar}197 to {dollar}-{dollar}183. Two identical clones were isolated. Northern blot analysis of Jurkat cell indicated the presence of a single band of approximately 2500 bases. The nucleotide sequence of the clones share a 99.5% similarity with a previously identified protein, namely DNA binding protein B (Dbpb).;Altogether, we have identified two transcription factors important in the regulation of IL-2 gene expression, namely NF-kB and RW-BF. The cDNA encoded protein has not yet been proven to be the protein that interacts with the {dollar}-{dollar}197 to {dollar}-{dollar}183 sequence in the Jurkat cells nuclear extracts, namely RW-BF.
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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.
