Research Department: Molecular Biology Graduation Date: May 2019
Abstract:The mammalian target of rapamycin complex 1 (mTORC1) is a master regulator for nutrient sensing, cell growth, and metabolism. mTORC1 is comprised of mTOR, Raptor, PRAS40, and mLST8. mTORC1 is hyperactivated in many human diseases including cancer, diabetes, and neurological disorders. Amino acids potently activate mTORC1 and promote mTORC1 lysosomal localization. Leu, Met, and Arg activate mTORC1 through the Rag GTPase pathway. In contrast, Gln promotes mTORC1 activation and lysosomal localization through ADP ribosylation factor 1 (Arf1). Because cancer cells are thought to be “addicted” to Gln, we are interested in identifying new components involved in Gln signaling to mTORC1. Using HAP1 Rag GTPase knockout (KO) cells we immunoprecipitated mTORC1 followed by mass spectroscopy (IP-MS), in order to identify novel mTORC1 interacting proteins. We followed up on 16 candidate proteins from IP-MS, by validating their interaction with mTORC1. 6 of the 16 proteins interacted with mTORC1. Moreover, we assessed the co-localization of mTORC1 with the 6 mTORC1 interacting proteins by confocal microscopy. The Gln signaling pathway is still highly uncharacterized, and the discovery of new components may lead to new targets for treating human disease such as cancer.
What does research mean to you? Conducting scientific research provides the unique opportunity to explore the unknown and constantly ask the question why. Regardless of what field the research is in, it amazes me to see the conclusions that can be drawn from simple techniques. I love being able to do with my own hands what I've read others do in textbooks. Rather then reading about bacterial transformations and ligations, it was cool for me to actually perform transformations and know the theory behind it. Tell us about your journey. During the weeks leading up to the Green Fellowship in the Spring, I was beyond ecstatic to be part of a well known research institution like UTSW. However, when I entered my lab, it was nothing like what I expected. Information was thrown at me left and right, and even though other lab members made themselves available for me to ask questions, I was expected to do the experiments on my own without anyone looking over my shoulder. Something I found out quickly trough having my own project is that the quality and quantity of your data will look like however much time and effort you put into it. At first, it was hard for me to adjust to working 40-50 hours a week and having to come in during weekends; however, by the end of the program I loved it. I enjoyed being the one responsible for my data, even when I sometimes got frustrated at experiments not working. By the end of the program, I had new techniques under my belt, and good data that I was proud of showing. I would have to say my favorite part of this research experience was seeing all my data finally come together in the form of a poster, and being able to interpret the data and see how it fit in with the overall research goals of my lab.
Advice for Future Green Fellows
Don't go into your UTSW lab and expect it to function the same as any of your past UTD labs. Each lab has a unique lab environment, so come in ready to adjust. Having your own project can be intense and time consuming, so try your hardest not to have any other commitments during your time in the program. Overall, enjoy your time at UTSW! Attend the weekly seminars, learn new techniques from your lab members, and read up on EVERYTHING you do in lab, whether it be understanding different techniques, looking up proteins you're working with, or staying abreast with any papers your lab has published.