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Title:Kinetics and thermodynamics of protein-RNA interactions and protein folding in vitro and in cells
Author(s):Guzman Sanchez, Irisbel
Director of Research:Gruebele, Martin
Doctoral Committee Chair(s):Martin Gruebele
Doctoral Committee Member(s):Gennis, Robert; Ha, Taekjip; Ceman, Stephanie S.
Department / Program:Biochemistry
Degree Granting Institution:University of Illinois at Urbana-Champaign
Subject(s):Protein-RNA interactions
protein folding
fluorescence resonance energy transfer (FRET)
fast relaxation imagining (FREI)
temperature jump
Abstract:Protein-RNA interactions and protein folding are critical subjects in biochemistry, because of their significance during the formation of active complexes and signaling pathways. Regardless of the substantial amount of studies in the fields of protein-RNA interactions and protein folding, little is known about the stability and kinetics of these in the cell. This doctoral dissertation aims to advance the understanding of protein-RNA interactions and protein folding inside cells through comparative in vitro studies, utilizing U1A-SL2 RNA complex and PGK/VlsE proteins as model systems, respectively. For the protein-RNA studies, dynamics experiments of one positive charged mutant of the spliceosomal U1A protein, the golden model for the RNA Recognition Motif (RRM), reveled a conformational transition for the protein only. Also, U1A-SL2 RNA dissociation kinetics studies with U1A positive charged mutants supported the previously proposed two-step dissociation pathway and demonstrated the importance of positive charge residues. The U1A-SL2 was also investigated in macromolecular crowded buffers were its binding affinity increased. It was also studied inside mammalian cells were it localized in the nucleus and its binding affinity decreased. For the protein folding studies, the extracellular VlsE antigen was found to be destabilized inside mammalian cells opposed to the intracellular PGK enzyme.
Issue Date:2015-04-20
Rights Information:2015 Irisbel Guzman Sanchez
Date Available in IDEALS:2015-07-22
Date Deposited:May 2015

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