Contribution of Muscleblind-Like1 (MBNL1) Binding Events to Dose-Response Behavior of MBNL1
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Date
2013-06
Authors
Gupta, Riti
Journal Title
Journal ISSN
Volume Title
Publisher
University of Oregon
Abstract
Mis-regulation of the alternative splicing factor muscleblind-like I (MBNL 1) plays a
significant role in the disease myotonic dystrophy (OM). MBNL 1 regulates alternative
splicing of genes involved in development of skeletal muscle, heart and the central
nervous system. In myotonic dystrophy the lack of properly localized MBNL 1 leads to
mis-splicing of many pre-mRNAs. One of the mis-spliced pre-mRNAs is the MBNLJ
pre-mRNA that codes for the MBNL 1 protein (an auto-regulated event). Specifically,
the mis-splicing is aberrant inclusion of exon 5 in the MBNLJ pre-mRNA. Previous
work has shown that intron 4 of the MBNLJ gene is highly conserved and contains
multiple MBNLI binding sites. It has been shown that a 90-nucleotide region within
intron 4 is necessary for regulation by MBNL 1. This study investigates the sensitivity of
the auto-regulated MBNLJ splicing event by generating MBNL 1 dose-response curves
using HEK293 cells with an inducible MBNL 1 expression system. Coupling this
expression system with MBNLJ constructs containing deletions of various MBNLI binding sites within the important 90 nucleotide regulation region, it was shown that a
single central MBNL1 site was fundamentally more important that other sites for
splicing regulation.
Description
39 pages. A thesis presented to the Department of Chemistry and the Clark Honors College of the University of Oregon in partial fulfillment of the requirements for degree of Bachelor of Science, Spring 2013.
Keywords
Muscleblind (MBNL1), RNA-binding proteins, Alternative splicing