Genome-wide Analysis of L(3)mbt and its Relationship to Drosophila melanogaster Insulators Open Access

Abstract

Lethal (3) malignant brain tumor [L(3)mbt] is a conserved transcriptional regulator. Chromatin
immunoprecipitation (ChIP) followed by high-throughput sequencing (ChIP-seq) analysis was
done to determine the genome-wide distribution of L(3)mbt in Drosophila melanogaster.
Results show that L(3)mbt is present at insulator sites in embryonic Kc cells; inverse levels of
enrichment of L(3)mbt and CP190 were also observed where binding sites overlap. Furthermore,
immunofluorescence microscopy of imaginal disc cells shows that L(3)mbt localizes to nuclear
bodies adjacent to CP190 insulator bodies. Comparison of L(3)mbt ChIP-seq data between two
cell lines revealed cell type-specific L(3)mbt enrichment sites, suggesting its role in differential
regulation of gene expression patterns.

Table of Contents

INTRODUCTION - 1

MATERIALS AND METHODS - 5

RESULTS

L(3)mbt is present at insulator sites - 8
L(3)mbt sequence motif corresponds to CP190 enriched motif - 11
L(3)mbt localizes to nuclear bodies near CP190 bodies - 12
Inverse enrichment levels of L(3)mbt and CP190 where binding sites overlap - 14
Functional analysis of L(3)mbt - 16

DISCUSSION - 22

REFERENCES - 24

LIST OF FIGURES AND TABLES

Figure 1 - Diagram of Drosophila insulators - 3
Figure 2 - Genome-wide localization of L(3)mbt - 9
Figure 3 - Distribution of L(3)mbt sites with insulators - 10
Figure 4 - Sequence logos - 11
Figure 5 - Immunofluorescence microscopy of imaginal disc cells - 13
Figure 6 - L(3)mbt enrichment correlates with lower CP190 levels - 14
Figure 7 - Analysis of l(3)mbt Kc-specific site - 19
Figure 8 - Analysis of l(3)mbt brain-specific site - 21

Table 1 - Gene ontology analysis of genes near L(3)mbt stable sites - 17
Table 2 - Gene ontology analysis of genes near L(3)mbt Kc-specific sites - 17
Table 3 - Gene ontology analysis of genes near L(3)mbt brain-specific sites - 17

About this Honors Thesis

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• Permission granted by the author to include this thesis or dissertation in this repository. All rights reserved by the author. Please contact the author for information regarding the reproduction and use of this thesis or dissertation.

School	Emory College
Department	Biology
Degree	BS
Submission	Honors Thesis
Language	English
Research Field	Biology, Genetics Biology, General
Keyword	CP190 Insulator Proteins ChIP-seq L(3)mbt
Committee Chair / Thesis Advisor	Corces, Victor, Emory University
Committee Members	Lucchesi, John, Emory University Weinschenk, Matthew, Emory University

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