Identification of Novel Protein-Protein Interactions in Primary Gingival Epithelial Cells

Dominika Burbul

Authors:  Dominika Burbul, Bernadett Papp

Faculty Mentor:  Bernadett Papp

College:  College of Dentistry

Abstract

When cells in the oral cavity undergo unfavorable alterations due to pathogen encounters, human diseases can arise. Therefore, we need a better understanding of the underlying molecular mechanisms to develop therapeutic interventions. A knowledge gap in the field arises from a lack of tools, limited access to human primary oral cells, and the loss of original cellular properties and protein interactions from experimentation. To overcome these barriers, my project aim is to create Yeast Two-Hybrid (Y2H) libraries from primary oral cells. We utilize Y2H since it is a well-established method to study protein-protein interactions and to identify novel protein interactions of target proteins. We have established two of these libraries, one from uninfected cells and the other from oncogenic Human Herpesvirus-8 (HHV-8) infected primary human oral cells. I am currently testing the quality of the two newly created libraries to make sure they contain a variety of expressed human genes in the oral cells. In the future, I hope to demonstrate the functionality of the new Y2H libraries by performing a proof-of-principle Y2H screen. The ultimate goal is to identify interaction partners of key oral factors and find out how they work together during host-pathogen interaction of oral cells.

Poster Pitch

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Poster

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Anna Walls
Anna Walls (@guest_1374)
1 year ago

What is the significance of a reporter gene?

Dominika Burbul
Dominika Burbul (@guest_1802)
Reply to  Anna Walls
1 year ago

Hey Anna!
A reporter gene is used to investigate a gene of interest. In my case, yeast colonies that turned blue signified the successful interaction between bait and prey proteins. It also signified the restoration of GAL4 activity. It can be an easy visual for the investigator to interpret.

Anna Walls
Anna Walls (@guest_1950)
Reply to  Dominika Burbul
1 year ago

Thank you!! You did a great job on on your poster by the way!

Chris Najjoum
Chris Najjoum (@guest_2180)
1 year ago

That was a very informative and well put together presentation! My question to you is what are the next steps you are going to take to after testing through DNA sequencing?

Dominika Burbul
Dominika Burbul (@guest_3628)
Reply to  Chris Najjoum
1 year ago

Hey Chris!
After doing steps 5 and 6 under the library screening heading, we hope to better understand what proteins are interacting. After doing this, we can hopefully link this to understanding the molecular mechanisms behind infection to create therapeutic interventions. We could always move further with the project by changing the bait protein we choose to interact with the prey proteins. There are lots of great ways to expand and continue the project!

Omaira Cheng
Omaira Cheng (@guest_2488)
1 year ago

Hi Dominika!
Great poster and presentation! I found the diagrams very helpful to understand the Y2H assay and screening procedure!

Dominika Burbul
Dominika Burbul (@guest_3650)
Reply to  Omaira Cheng
1 year ago

Thank you so much!

ZT
ZT (@guest_2752)
1 year ago

Hi Dominika, this is a nicely organized poster. Can you tell how much time it takes to see the blue colonies indicating protein-protein interaction after the yeast plasmids have been transformed into yeast? Tx

Dominika Burbul
Dominika Burbul (@guest_3980)
Reply to  ZT
1 year ago

Hello!
Thank you for the nice comment! I believe the blue colonies appeared after just a few days, about three, indicating their protein-protein interactions.

Bernadett Papp
Bernadett Papp (@guest_2864)
1 year ago

Hi Dominika!

Good job presenting your USP research project!

You are establishing a new method to identify novel human protein-protein interactions of oral cells.

It is not easy to develop new tools for research. It takes time, but it is always worth the effort, when we innovate!

Can you tell us, how long it takes for the yeast colonies to become visible on the plates, and to turn blue in your experience? Do you have to wait a long time? What is the ideal temperature for the yeast to grow?

Good job! Thanks! (BP)

Dominika Burbul
Dominika Burbul (@guest_4278)
Reply to  Bernadett Papp
1 year ago

Hello Dr. Papp!
It only took around three days for yeast colonies to become visible on the plates. The ideal temperature for the yeast to grow was 30 degrees C. Regarding both of these questions, there is a range you could play with to achieve optimum results.
Thank you for visiting my page!

Gabriela Peguero Kushner
Gabriela Peguero Kushner (@guest_5840)
1 year ago

Great job! Really nice and easy to follow poster. I was just curious about what the HA is in the plasmid is, what is their purpose?

Thanks!

Gabriela Peguero