INVESTIGATING THE COMPETITION BETWEEN COMPONENTS OF DUAL-FUNCTION RNA

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dc.contributor.advisorStorz, Gisela Ten_US
dc.contributor.authorAoyama, Jordan James Masuoen_US
dc.contributor.departmentMolecular and Cell Biologyen_US
dc.contributor.publisherDigital Repository at the University of Marylanden_US
dc.contributor.publisherUniversity of Maryland (College Park, Md.)en_US
dc.date.accessioned2021-07-13T05:33:52Z
dc.date.available2021-07-13T05:33:52Z
dc.date.issued2021en_US
dc.description.abstractNon-coding RNAs (ncRNAs) and small proteins have both emerged as important regulators of gene expression. Dual-function RNAs encode a small protein and have a separate function as a regulatory RNA. Although first discovered in bacteria, dual-function RNAs have now been identified and characterized in eukaryotes as well. These RNAs allow two activities of a single gene to regulate targets at multiple levels. The work described here explores how two novel and one synthetic dual-function RNA act and how competition between the components of a dual-function RNA impacts their functions. AzuCR is a 164-nucleotide E. coli RNA that was previously shown to encode a 28 amino acid protein (AzuC). This work demonstrates that the AzuC small protein impacts glycerol metabolism, with the small protein increasing activity of GlpD, an essential enzyme in glycerol catabolism, while the RNA base pairs with and represses galE mRNA, a gene essential for galactose metabolism. The second dual-function RNA studied in this work is Spot 42, a 109-nucleotide RNA known to base pair with and repress mRNAs encoding proteins involved in the metabolism of non-preferred carbon sources. Although Spot 42 is a well-characterized base pairing small RNA (sRNA) in E. coli, this work shows it also encodes a 15-amino acid protein, SpfP. SpfP was found to bind to cAMP receptor protein (CRP) and block activation of some target genes. For both AzuCR and Spot 42, the coding sequence for the small protein overlaps the base pairing region, and we have observed that translation interferes with base pairing activity suggesting competition between the sRNA and mRNA activities. Finally, a synthetic dual-function RNA was constructed from the Escherichia coli sRNA MgrR and the mRNA for the small protein MgtS. Various versions of this hybrid molecule are used to probe how the organization of components is important for the proper functioning of a dual-function RNA. These three studies highlight the complexities of regulation by dual-function RNAs and provide insights into how these molecules coordinate two different activities to carry out regulatory roles in the cell.en_US
dc.identifierhttps://doi.org/10.13016/9zex-c2s8
dc.identifier.urihttp://hdl.handle.net/1903/27363
dc.language.isoenen_US
dc.subject.pqcontrolledMicrobiologyen_US
dc.subject.pqcontrolledBiochemistryen_US
dc.subject.pquncontrolledDual-function RNAen_US
dc.subject.pquncontrolledregulatory RNAen_US
dc.subject.pquncontrolledSmall proteinsen_US
dc.subject.pquncontrolledsmall RNAen_US
dc.titleINVESTIGATING THE COMPETITION BETWEEN COMPONENTS OF DUAL-FUNCTION RNAen_US
dc.typeDissertationen_US

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