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Date Permissions Signed

10-7-2012

Date of Award

2012

Document Type

Masters Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Pillitteri, Lynn

Second Advisor

Otto, Joann

Third Advisor

Rose, Jacqueline K.

Abstract

Proper development of neuronal circuits are crucial for nervous system functioning. A novel pathway regulating axon and synapse development in Caenorhabditis elegans through nuclear 3'-end polyadenylation of nascent mRNA has recently been uncovered (Van Epps et al., 2010). In this pathway, the protein product of the gene synaptic defective enhancer (sydn-1) negatively regulates polyadenylation factor subunit homolog (PFS-2), an evolutionarily conserved scaffolding protein in a multi-protein complex involved in mRNA 3' -end processing. Although 3'-end processing of mRNA has a regulatory role in many cellular processes, regulation of synapse and axon development via this cellular mechanism has not been characterized. An RNAi screen was performed using C. elegans to identify genetic suppressors in this regulatory pathway. This screen was conducted in a sensitized genetic background that allowed for targeted gene silencing and visual screening for suppression of a neuronal phenotype resulting from SYDN-1 mediated misregulation of PFS-2. We identified seven novel genetic interactors. One of these genes, ELAV-type RNA binding protein family (etr-1), encodes a highly conserved protein that is involved in 3'-end alternative splicing during muscle development. The human homolog of ETR-1, CUGBP, elav-like family member (CELF1), is implicated in the development of the genetic disease myotonic dystrophy. Vectors were produced to further study the function of these proteins in the context of the SYDN-1/PFS-2 pathway. One vector contains etr-1 cDNA fused to a gene encoding GFP, and the other contains CELF1 cDNA fused to the yeast-two-hybrid GAL4 DNA-binding domain, to be used for in vivo localization and yeast-two-hybrid assays, respectively.

Type

Text

DOI

https://doi.org/10.25710/yye4-2a12

Publisher

Western Washington University

OCLC Number

818256557

Subject – LCSH

Caenorhabditis elegans--Genetics; Genetic regulation; Gene silencing; RNA; Neural circuitry

Format

application/pdf

Genre/Form

masters theses

Language

English

Rights

Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this thesis for commercial purposes, or for financial gain, shall not be allowed without the author's written permission.

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