Type
Text
Type
Thesis
Advisor
Smith, Steven. | Czaplinski, Kevin
Date
2013-12-01
Keywords
EMSA, Hnrnpab, NMR, RRM | Biochemistry
Department
Department of Biochemistry and Cell Biology.
Language
en_US
Source
This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.
Identifier
http://hdl.handle.net/11401/76905
Publisher
The Graduate School, Stony Brook University: Stony Brook, NY.
Format
application/pdf
Abstract
Post-transcriptional trafficking of mRNA within the cell provides for spatial regulation of protein expression. Cis-acting localization elements known as zip codes located often in 3' untranslated regions (UTRs) of mRNA transcripts play a strong role in directing distribution within the cell through recognition by RNA-binding proteins. Hnrnpab is a protein with putative roles in this process with RNA-binding properties likely directed through tandem RNA Recognition Motifs (RRMs) that form a cooperative RNA Binding Domain (RBD). We purified this Hnrnpab RBD and carried out electrophoretic mobility shift assays (EMSAs) using the zip code region from the ActB gene to demonstrate high specificity binding. In order to determine the structural determinants of this specificity we purified and 13C, 15N-labeled Hnrnpab RBD for analysis by solution NMR to begin determining a structure for the RRMs both with and without high specificity RNA partners. Further characterization of how Hnrnpab recognizes its targets and what structural motifs determine this type of binding will facilitate greater understanding of the spatial regulation of protein products necessary for cellular and organismal development. | 34 pages
Recommended Citation
Catt, Daniel, "Structural and Biochemical Analysis of the Hnrnpab Tandem RRM RNA Binding Domain" (2013). Stony Brook Theses and Dissertations Collection, 2006-2020 (closed to submissions). 2779.
https://commons.library.stonybrook.edu/stony-brook-theses-and-dissertations-collection/2779