Date of Award
Spring 5-12-2022
Degree Type
Thesis
Degree Name
Master of Science in Integrative Biology (MSIB)
Department
Biology
Committee Chair/First Advisor
Scott Nowak
Major Professor
Scott J. Nowak
Second Committee Member
Jennifer Cooper
Third Committee Member
Christopher Cornelison
Abstract
Akirin is a small highly conserved nuclear protein that has been of scientific interest since its discovery in 2008. Akirin functions as a transcriptional co-factor for many important biological pathways; however, detail regarding Akirin’s mechanism remains unclear. Of the 4 major animal chromatin remodeling complexes, SWI/SNF, INO80, ISWI, and CHD, we know Akirin interacts with subunits of the SWI/SNF family Brahma (BRM) chromatin remodeling complex during myogenesis and cardiogenesis. Interaction between Akirin and CHD family chromatin remodeling complexes has not been determined.
Our study aimed to examine the colocalization of Akirin with the CHD family chromatin remodeling complex known as the nucleosome remodeling and deacetylase (NuRD) complex. Drosophila polytene chromosomes were used to visually identify colocalization between Akirin and subunits of the NuRD complex. The NuRD subunit, HDAC1, functions to remove acetyl groups from histone tails. Therefore, the colocalization between Akirin and histones modified by acetylation and methylation were also examined.
A novel polytene squash method was developed as a byproduct of our study to improve the quantity of usable images for analysis. The vibrating tip of a regrout tool was adapted with a rubber tip and applied to the coverslip of microscope slides containing polytene chromosomes to aid in gently tensing apart chromosome arms.
The NuRD complex subunits, Mi-2 and HDAC1, showed remarkable colocalization with Akirin along Drosophila polytene chromosome arms. Also, Akirin was found to be largely omitted from regions containing acetyl histone modifications.