File
Molecular characterization of the coding region determinant-binding protein interaction with KRAS oncogene mRNA
Digital Document
Abstract |
Abstract
The coding region determinant binding protein (CRD-BP) plays a role in some cancers, regulating gene expression through mRNA trafficking, turnover, and translational repression. Recent evidence reveals CRD-BP binding to KRAS mRNA '" a proto-oncogene frequently mutated and/or overexpressed in cancer, leading to up-regulation. A CRD-BP-KRAS mRNA interaction therefore presents a novel therapeutic opportunity to target KRAS-driven cancers and is the aim of this thesis. Electrophoretic mobility and immunoprecipitation experiments presented here using GXXG-mutant CRD-BP variants show four K-homology domains are required to bind a 57-nucleotide minimum binding sequence in the coding region of KRAS RNA these mutations do not compromise protein folding as evidenced by circular dichroism spectroscopy. DNA-based anti-sense oligonucleotide inhibitors were developed based on the minimum binding sequence that successfully abrogate the CRD-BP-KRAS mRNA interaction in vitro. Finally, a CRD-BP KH1toKH4 construct purified using a novel '~on-column'-refolding scheme suggests a possible role for RRM-domains as negative regulators of RNA-binding. --Leaf i. |
---|---|
Persons |
Persons
Author (aut): Mackedenski, Sebastian J.
Thesis advisor (ths): Lee, Chow
|
Degree Name |
Degree Name
|
Department |
Department
|
DOI |
DOI
https://doi.org/10.24124/2015/bpgub1098
|
Collection(s) |
Collection(s)
|
Origin Information |
|
||||||
---|---|---|---|---|---|---|---|
Organizations |
Degree granting institution (dgg): University of Northern British Columbia
|
||||||
Degree Level |
Subject Topic |
Subject Topic
|
---|---|
Library of Congress Classification |
Library of Congress Classification
RC268.4 .M65 2015
|
Extent |
Extent
Number of pages in document: 137
|
---|---|
Physical Form |
Physical Form
|
Content type |
Content type
|
Resource Type |
Resource Type
|
Genre |
Genre
|
Language |
Language
|
Handle |
Handle
Handle placeholder
|
---|
Use and Reproduction |
Use and Reproduction
Copyright retained by the author.
|
---|---|
Rights Statement |
Rights Statement
|
Download
Language |
English
|
---|---|
Name |
Molecular characterization of the coding region determinant-binding protein interaction with KRAS oncogene mRNA
|
Authored on |
|
MIME type |
application/pdf
|
File size |
16143835
|
Media Use |