Researcher ORCID Identifier

0000-0003-2327-9852

Graduation Year

2023

Date of Submission

12-2022

Document Type

Campus Only Senior Thesis

Degree Name

Bachelor of Arts

Department

Biology

Reader 1

Aaron Leconte

Reader 2

Ryan Boileau

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Terms of Use for work posted in Scholarship@Claremont.

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© 2022 Abrahan Vasquez

Abstract

The epithelial-mesenchymal transition (EMT) is relevant to cancer studies since it is responsible for cell migration, an essential component of metastasis. In examining the epigenetics of cell transitions such as the EMT, it is important to understand the interactions between different non-coding regions of DNA due to their gene regulatory effects. Specifically, investigating enhancer activation allows for an understanding of three-dimensional enhancer-promoter interactions. The transcription factor Grainyhead-like 2 (GRHL2) can suppress the EMT in certain cell types and is involved in bringing together enhancers and promoters through recruitment of the cofactor protein cohesin. Here, we use a GRHL2-Estrogen fusion construct that allows for a reductionist model of the transition from naive embryonic stem cells to formative epiblast-like cells, a transition relevant to the EMT since it is an in vitro substitute for early cell fate differentiation, and it features GRHL2 enhancer-switching. This reductionist system has allowed for an investigation of the transcriptional requirement of the coactivator protein subunits, RAD21, BRG1, and CBP/p300. Preliminary data from the Blelloch lab suggests that some of these coactivators, namely RAD21, may not be required for transcription, and a time-course degradation of these coactivators and qPCR RNA level analysis of CLDN6 and WNT7B, two direct downstream transcriptional targets of GRHL2, further confirms the dispensability of RAD21 but suggests further optimizing of the model system for the other coactivators. With the requirement of RAD21 now in question, future research will center on optimization of the model and on resolving the order of coactivator activity.

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