Interacting with Proteins to Find Protein Interactions

Author: Elise Wyatt       Major: Biomedical Engineering 

Creating Cell Cultures

On the first day of my cell biology class spring of my sophomore year, my soon-to-be research mentor, Dr. Yuchun Du, was introducing himself and his research. It was through his presentation that I became interested in this lab and soon joined his team. For the first semester, I worked closely with the senior research associate, Jianhong Zhou, who taught me different lab techniques that I would need for my project. I was able to gain hands-on experience in checking protein concentrations, creating gels, developing Western blots, and more. After studying abroad the following semester, I was back in Dr. Du’s lab ready to start my project. One of the many projects Dr. Du’s lab is working on is the influenza A virus (IAV), which is responsible for seasonal flu outbreaks. My project was to find which domain of the cellular host protein DEAD-box protein 1 (DDX1) interacts with the IAV nonstructural protein-1 (NS1). This interaction is thought to play an important role in the replication of IAVs and thus elucidating which domain of DDX1 that binds NS1 could be useful in understanding viral mechanisms for establishing a successful host infection.

In order to determine the domain of DDX1 responsible for the interaction with NS1, an in vitro protein binding assay with His-tagged DDX1 fragments and GST-tagged NS1 protein was performed and then examined using Western Blotting against His-tag and GST-tag. From this, it was determined that domain I of DDX1 interacts with NS1.

For this project, the Ph.D. candidate, Brihget Sicairos, helped me immensely. She has been working with Dr. Du’s lab for a few years and taught me the science behind the experiments and what was happening on a microscale. Ms. Zhou and Ms. Sicairos worked hard on my project when the pandemic caused the University of Arkansas to go remote. It is unfortunate that I was unable to work in the lab from March to August in 2020, however, thanks to their dedication I was able to finish my project.

My mentor was crucial for the success of my thesis and defense. He is very knowledgeable in writing research papers and it was a privilege to have his insight on my work. Through his comments and suggestions, I was able to learn how to properly write a scientific paper and create a successful thesis. I am very thankful he gave me the opportunity to work in his lab because I was able to gain valuable skills which will help me in my future career.

Beginning fall 2021, I will be attending the University of Oklahoma College of Medicine where I will pursue my Doctor of Medicine (MD). While there, I plan on getting involved in a research lab focused on immuno-oncology. The Honors College Research Grant allowed me to continue my research of the influenza A virus and how it interacts with host cellular protein DDX1. Through my time in Dr. Du’s lab, I was able to apply what I was learning in my biomedical engineering curriculum to my work which helped me gain a better understanding of recombinant technology, SDS-PAGE, Western blotting, and much more. I am grateful for my time in Dr. Du’s lab and for all the support I received from his team and the Honors College and I am excited to start the next chapter of my academics.