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

Want, Yiwen (Spring 2021)

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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

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