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Title:Biochemical and genetic characterization of U1 and U3snRNAs from Schizosaccharomyces pombe
Author(s):Porter, Gregory Lewis
Doctoral Committee Chair(s):Wise, Jo Ann
Department / Program:Biology, Molecular
Biology, Genetics
Biology, Microbiology
Chemistry, Biochemistry
Discipline:Biology, Molecular
Biology, Genetics
Biology, Microbiology
Chemistry, Biochemistry
Degree Granting Institution:University of Illinois at Urbana-Champaign
Subject(s):Biology, Molecular
Biology, Genetics
Biology, Microbiology
Chemistry, Biochemistry
Abstract:I have examined the small RNA profile of the fission yeast Schizosaccharomyces pombe. In vivo labeling followed by two dimensional gel electrophoresis of S. pombe RNA demonstrates the presence of four prominent species in the size range of interest (100-300 nucleotides). Further analysis of these has shown that they represent U1, U2, U3, and 7SL homologs.
I have cloned, sequenced, and disrupted the gene encoding U1 small nuclear RNA in Schizosaccharomyces pombe. This RNA exhibits a high degree of secondary structure homology to human U1. S. pombe U1 lacks two nucleotides just following the 5$\sp\prime$ cap structure which are present in all other U1 homologs examined to date, and the region which corresponds to the binding site for the human 70K protein is more divergent than in other organisms. A putative upstream transcription signal is conserved in sequence and location among all loci encoding spliceosomal snRNAs in S. pombe with the exception of U6. Disruption of the single copy U1 gene, designated snu1, reveals that this RNA is indispensable for viability. I have constructed mutants in the 5$\sp\prime$ region of the U1 RNA.
I have cloned and sequenced the two genes encoding a 255 nucleotide small nuclear RNA from the fission yeast Schizosaccharomyces pombe. Based on the presence of four regions of primary sequence conservation and a predicted secondary structure similar to that previously proposed for human U3, I conclude that this molecule is the fission yeast homologue of this mammalian snRNA. I propose revised secondary structures containing two hairpins for this portion of the U3-like snRNAs from Schizosaccharomyces pombe, Saccharomyces cerevisiae and Dictystelium discoideum.
I have developed a hypothesis which states that the 3$\sp\prime$ splice sites of some introns are recognized by base-pairing to nucleotides C$\sb9$U$\sb{10}$ of U1 snRNA. Evidence and implications are discussed.
Issue Date:1990
Rights Information:Copyright 1990 Porter, Gregory Lewis
Date Available in IDEALS:2011-05-07
Identifier in Online Catalog:AAI9114376
OCLC Identifier:(UMI)AAI9114376

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