Identification of novel histone locus body components in Drosophila melanogaster Open Access

Abstract

The conserved histone locus body (HLB) facilitates replication-dependent histone biogenesis during the early developmental stages in most eukaryotic organisms. However, many regulatory factors at the HLB have not been identified. In this paper, I use two research approaches, an unbiased proteomic screening method, and a bioinformatics-based protein candidate method, to identify potential protein factors that target the histone genes. For the unbiased proteomic screening, I adopted a dCas9 (inactive Cas9)-gRNA sequence-specific system that targets the histone locus in D. melanogaster. Future work with this system will discover new HLB factors by mass spectrometry. For the bioinformatics-based candidate approach, I mapped publicly available ChIP-seq datasets of potential HLB factors to a single copy of the histone gene array of Drosophila melanogaster. I identified SIN 187 and SIN 220, two major isoforms of the SIN3 complex, as candidates that target the histone locus. I expect to test my hypothesis through subsequent immunostaining experiments. Overall, through the two mentioned experimental approaches, I expect to contribute to the further understanding of HLB regulatory factors at the histone locus and their contribution to the coordinated histone biogenesis in Drosophila melanogaster.

Table of Contents

Chapter 1: Introduction

Chapter 2: Identification of novel HLB candidates by dCas9-targeted locus specific protein isolation in Drosophila melanogaster

Chapter 3: Identification of SIN3 as an HLB candidate by a bioinformatics-based protein candidate approach

Chapter 4: Mapping of ChIP-seq data in Galaxy fails to show localization of the gypsy insulator complex or the M1BP insulator to the histone array

Chapter 5: Discussions and Future Directions

About this Honors Thesis

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School	Emory College
Department	Biology
Degree	B.S.
Submission	Honors Thesis
Language	English
Research Field	Biology, Genetics
Keyword	Histone locus body Histone biogenesis Protein identification method Histone gene transcription Bioinformatics Drosophila melanogaster Galaxy dCas9-gRNA
Committee Chair / Thesis Advisor	Leila Rieder, Emory University
Committee Members	Judith Fridovich-Keil, Emory University Arri Eisen, Emory University

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