Interactions between domino and other chromatin protein encoding loci Open Access

Abstract

The Notch signaling pathway plays an integral role in cell signaling. Our lab previously determined genetic interactions between Drosophila Notch and a co-activator known as Mastermind (Mam). Our lab also screened for and identified interactions between Mam and transposon-generated random genomic insertions. The screen found loss of function (LOF) domino, which encodes a chromatin remodeling protein, strongly enhanced the Mam wing phenotype. The lab then constructed a recombinant chromosome containing a Gal4 driver and UAS driven RNAi constructs directed against domino (C96-domR). This chromosome leads to a dominant, partially-penetrant wing nicking phenotype. Domino plays a role in hematopoiesis, cell growth and proliferation, apoptosis, histone exchange during DNA repair, maintenance of stem cells, and autophagy. In this study, we focus on the wider gene network of domino. We attempt a rational approach, by testing other chromatin protein encoding loci identified by Van Bemmel et al. (2013). With the use of the DNA adenine methyltransferase identification (DamID) system, Van Bemmel et al. (2013) identified 42 proteins that had yet to be associated with chromatin, and 70 previously known chromatin proteins. We performed a genetic screen using RNAi against 30 of the Van Bemmel loci for modifiers of LOF domino. Using a wing-nicking assay, we found 25/30 Van Bemmel loci significantly enhanced or suppressed the C96-domR phenotype. Additionally, we found a subset of Van Bemmel loci genetically interact with GOF domino. Also, several of the Van Bemmel loci modify LOF domino in the eye. Since our lab recently found LOF domino leads to cell death along the wing margin, we investigated whether the modification in the C96-domR wing nicking phenotype was due to a change in cell death. We found that RNAi against Van Bemmel locus, Caf-1, synergistically increases cell death with LOF domino. The present study validates C96-domR is an excellent tool for assaying chromatin function. Further, since the majority of the modifiers in our screen are associated with active chromatin, our study illuminates Domino's role as a gene activator. We confirm that Domino is a protein with pleiotropic functions and widen the existing understanding of Domino's interaction with other chromatin-associated proteins.

Table of Contents

INTRODUCTION. 1

FIGURE 1. 5

MATERIALS AND METHODS. 13

FIGURE 2. 14

RESULTS. 16

FIGURE 3. 17

FIGURE 4. 18

TABLE 1. 19

TABLE 2. 19

TABLE 3. 20

TABLE 4. 20

TABLE 5. 21

TABLE 6. 21

TABLE 7. 22

TABLE 8. 23

TABLE 9. 25

FIGURE 5. 26

DISCUSSION. 27

TABLE 10. 36

REFERENCES. 42

About this Honors Thesis

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School	Emory College
Department	Biology
Degree	BS
Submission	Honors Thesis
Language	English
Research Field	Biology, Genetics Biology, General
Keyword	Drosophila Notch Domino chromatin cell death
Committee Chair / Thesis Advisor	Yedvobnick, Barry, Emory University
Committee Members	Deal, Roger, Emory University Katz, David J, Emory University

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