Friday, November 8, 2013

My Dissertation (Intracellular trafficking of BK Virus: from the ER to the nucleus)



So, I set a date to do my thesis defense, on December 13. This means I have until the end of November to submit my dissertation to my committee… which means, I should be working on it right now. Instead, I thought I would try to explain my work here where I don’t have to use big words all the time.

The BK Virus (Signficance)
The lab that I work in does research on BK Polyomavirus. The BK does not stand for anything. Would it really matter if it did? Call it the Burger King virus like my family does. Anyway, “BK” is a virus that is actually already inside 90% of people because we pick it up when we are dirty kids. But that doesn’t matter, because healthy people aren’t really affected by the virus and it just hangs out quietly throughout your life. That means it’s actually very good at being a virus because it is able to quietly spread through the population without making a mess… that’s a conversation for another time, though.
Why do we study this virus? We get funding to study this virus because it can cause problems for people on immunosuppression, especially people with kidney transplants. When the immune system is taken out of the picture, the virus gets out of control replicating in the kidney cells and can basically destroy the new kidney. 
Importantly, we don’t have any anti-viral treatments for BK. This is partly because there a lot things we do not know about how the virus works. That’s where I come in! And other researchers.

My Research (Introduction)
The way that anti-virals work is they usually block a process in the virus life cycle.  There are a lot of different things that need to be known about cells and the virus and how they interact if you are going to make a drug that gets in the way. So, basically my goal is to understand how the BK virus interacts with the cells. I am doing that by figuring out how it travels (or “traffics”) through the cell. I am not looking specifically for a cure for BK infection, but I am learning all about how the virus uses and messes up normal cell biology. Then, someone else can use that information to make informed decisions about what drugs might stop BK.

How do we figure this stuff out? (Materials and Methods)
Basically, I work with human kidney cells in a petri dish, and I infect them with virus. All you have to do is put some pure virus on top of the cells, they will become infected. I can tell they are infected by looking for a certain virus protein after a day or two. So, to answer my question of what the virus interacts with inside the cell, the main way to do that is to take away certain things in the cell, and see whether they can still be infected, and what is happening if they can’t be infected. Interestingly, the cell can still function pretty well without specific proteins or pathways.
We can take away cellular functions in a few different ways. The method I’ve used most often is I treat the cells with a drug that specifically stops certain proteins and cellular machinery from working. These types of drugs are often only used in research to help figure things out. They are super convenient.

What did I find? (Results)
Do you remember high school cell biology? What I found is that BK travels from the outside of the cell, into the endoplasmic reticulum (the ER), then into the cytosol by using something call the proteasome and ER-associated degradation pathway, and gets into the nucleus by using the nuclear pore. I’ve illustrated it below.
At this point you might be like, gosh who cares?! This is boring. Well, shut up. Its super cool that a virus can go from outside your body to inside your body and then inside your cells and then travel through the cell and disassemble and put its own DNA into the nucleus of your cells and replicate. Watch the Battlefield Cell video!

What does all this work I did contribute to science? (Discussion)
The goal of scientific research is obviously to contribute to science by providing information that was not known before. My research hasn’t been super ground-breaking, because we had hypothesized that these things were true based on work that had been done with similar viruses, and then gave evidence that they were true. But that’s what a lot of science involves, just supporting hypotheses that are out there but haven’t been given much "convincing" evidence yet. My hard work has contributed to the field of virus research, whether it was earth-shattering or not, because it has provided data that did not exist before.

What helped me finish grad school? (Acknowledgments)
I would like to acknowledge these things that have been with me through these past five years:
anti-anxiety meds, alcohol, cardio workouts, cookies, things that remind me why I love science, beer, friends that make me laugh, realizing that I can still be called a scientist after failing, people who tell me I’m smart.