Type

Text

Type

Thesis

Advisor

Gergen, John P | Erezyilmaz, Deniz F

Date

2017-12-01

Keywords

Biochemistry | Cytology

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/11401/78220

Publisher

The Graduate School, Stony Brook University: Stony Brook, NY.

Format

application/pdf

Abstract

The slp1 gene has two upstream enhancer cis-regulatory modules that regulate transcription by non-additive interactions, the Proximal Early Stripe Element (PESE) and the Distal Early Stripe Element (DESE). These two enhancers are regulated by pair-rule transcription factors Even-skipped (Eve), Odd-Pared (Opa), Runt, and Fushi tarazu (Ftz). Both enhancers are repressed by Runt and Ftz, but only PESE is repressed by Eve. PESE and DESE are regulated along the antero-posterior axis of the 3hr Drosophila embryo while the short-gastrulation (sog) enhancer responds to regulation on the dorso-ventral axis. By combining the sog enhancer in a reporter gene construct with the PESE enhancer and the slp1 p381 promoter, it is possible to investigate whether these heterologous enhancers can interact in a non-additive manner. Previous in-situ hybridization results suggest that the sog enhancer can facilitate transcription at the slp1 transcription start site (TSS). When Chromatin Immunoprecipitation (ChIP) was preformed on 3hr embryos with ectopically expressed Runt and Ftz, qPCR results showed an increase of RNA polymerase II (Pol II) at the PESE-sog p381 reporter gene promoter. These results suggest sog enhancer is able to overcome the repression mediated by PESE and interact with the pre-initiation complex machinery at the slp1 promoter. | 35 pages

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