LE3 .A278 2019

2019

Easy, Russell
Langelaan, David N.

Acadia University

Bachelor of Science

Honours

Biology

Biology

Human skin is an important barrier against pathogens and excessive water loss. In the skin, specialized cells known as melanocytes synthesize melanin, which provides protection from damage caused by ultraviolet radiation, which can lead to skin cancer. During melanocyte development, several transcription factors are critical to control gene expression and coordinate cellular growth and differentiation. In particular the Microphthalmia-associated transcription factor (MITF) is the ‘master regulator’ that controls melanocyte differentiation and melanin synthesis. MITF continues to influence melanoma properties and is essential for cellular proliferation. To control cellular properties, MITF binds to gene promotors using a central basic-helix-loop-helix DNA binding motif. The N-terminal and C-terminal regions of MITF then interact with the transcriptional co-activators CREBs binding protein (CBP) and p300 (CBP/p300), which acetylate histones and activate gene transcription. Despite the influence that MITF has on melanoma, there is very little structural or mechanistic knowledge of how MITF functions. Using a combination of protein interaction assays, nuclear magnetic resonance spectroscopy, and biophysical techniques, we determined that the N-terminal activation domain of MITF binds with high affinity to the TAZ2 protein domain of CBP/p300. These results will help to determine how MITF interacts with transcriptional co-activators to regulate gene transcription and will provide strategies to disrupt MITF function for the potential treatment of melanoma.

The author retains copyright in this thesis. Any substantial copying or any other actions that exceed fair dealing or other exceptions in the Copyright Act require the permission of the author.

https://scholar.acadiau.ca/islandora/object/theses:3276