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Adenovirus Strategies for Altering the Cellular Environment in Favor of Infection

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Adenovirus Strategies for Altering the Cellular Environment in Favor of Infection University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2019 Adenovirus Strategies For Altering The Cellular Environment In Favor Of Infection Christin Herrmann University of Pennsylvania Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Allergy and Immunology Commons, Immunology and Infectious Disease Commons, Medical Immunology Commons, and the Virology Commons Recommended Citation Herrmann, Christin, "Adenovirus Strategies For Altering The Cellular Environment In Favor Of Infection" (2019). Publicly Accessible Penn Dissertations. 3568. https://repository.upenn.edu/edissertations/3568 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/3568 For more information, please contact [email protected]. Adenovirus Strategies For Altering The Cellular Environment In Favor Of Infection Abstract Viruses, as obligate intracellular pathogens, rely on their host cell for successful replication. Viruses have evolved different strategies to hijack and redirect cellular processes to benefit infection and overcome host immune responses. Understanding the mechanisms by which viruses exploit their host cells will reveal new targets for antiviral therapies. In addition, these studies can provide insights into the regulation of fundamental cellular processes. While much progress has been made in this area, many unexpected nuances of virus-host interaction are still being discovered. Here, we employed several strategies to uncover new aspects of viral manipulation of the host environment by adenovirus, a nuclear-replicating DNA virus that commonly infects humans. The first project focused on how viral histone-like proteins impact cellular chromatin. Adenovirus encodes the small, basic protein VII that coats and condenses viral genomes. The effect of this viral DNA-binding protein on host chromatin structure and function had remained unexplored. Here we demonstrated that protein VII interacts with host nucleosomes and is sufficiento t alter nuclear morphology. We also identified post-translational modifications of protein VII that regulate chromatin association. Through a proteomics analysis of chromatin composition, we revealed that protein VII causes nuclear retention of HMGB1, a host alarmin, and reduces downstream inflammation. The second project examined roles of viral-mediated ubiquitination during infection. Ubiquitination of host proteins, mediated by adenovirus proteins E1B55K and E4orf6, is important for viral RNA processing. However, previously identified substrates of viral-mediated ubiquitination do not explain this phenotype. Here we used a proteomics approach to define new substrates of the E1B55K/E4orf6 complex. We uncovered viral-mediated ubiquitination of RNA-binding proteins (RBPs) which, unlike ubiquitination of other substrates, does not result in proteasomal degradation. We furthermore demonstrated that ubiquitination of RBPs RALY and hnRNP-C decreases their binding to viral RNA and relieves a restriction these host proteins exert on adenovirus RNA processing. Our study of adenovirus proteins revealed new strategies employed by viruses to alter host functions: manipulating host chromatin through viral histone-like proteins to dampen immune responses and regulating RNA processing by non-degradative ubiquitination of cellular RBPs. Degree Type Dissertation Degree Name Doctor of Philosophy (PhD) Graduate Group Cell & Molecular Biology First Advisor Matthew D. Weitzman Second Advisor Carolina B. Lopez Keywords Adenovirus, Chromatin, RNA, Ubiquitin, Virus Subject Categories Allergy and Immunology | Immunology and Infectious Disease | Medical Immunology | Microbiology | Virology This dissertation is available at ScholarlyCommons: https://repository.upenn.edu/edissertations/3568 ADENOVIRUS STRATEGIES FOR ALTERING THE CELLULAR ENVIRONMENT IN FAVOR OF INFECTION Christin Herrmann A DISSERTATION in Cell and Molecular Biology Presented to the Faculties of the University of Pennsylvania in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy 2019 Supervisor of Dissertation _____________________ Matthew D. Weitzman, Ph.D. Professor of Pathology and Laboratory Medicine Graduate Group Chairperson _______________________ Daniel S. Kessler, Ph.D. Associate Professor of Cell and Developmental Biology Dissertation Committee Carolina B. Lopez, Ph.D., Associate Professor of Microbiology and Immunology Sara Cherry, Ph.D., Professor of Microbiology Paul M. Lieberman, Ph.D., Professor of Gene Expression and Regulation Paul F. Bates, Ph.D., Professor of Microbiology ADENOVIRUS STRATEGIES FOR ALTERING THE CELLULAR ENVIRONMENT IN FAVOR OF INFECTION COPYRIGHT 2019 Christin Herrmann This work is licensed under the Creative Commons Attribution- NonCommercial-ShareAlike 4.0 License To view a copy of this license, visit https://creativecommons.org/licenses/by-nc-sa/4.0/us/ DEDICATION Ich widme diese Arbeit meinen Eltern, die mich immer darin unterstützt haben meinen eigenen Weg zu gehen. iii ACKNOWLEDGMENTS I want to thank everyone that has contributed to this great experience of curiosity, inventiveness, learning, many mistakes, and exhilarating successes that is grad school and science in general. I am exceedingly grateful to all of you for your support! I especially want to thank my advisor Matt. His mentorship, high expectations, and unwavering support have helped me become a better scientist. Matt has taught me the importance of asking the right questions and to always have high expectations of myself. His guidance has enabled me to successfully drive my own research projects, making me ready to face the challenges that are waiting for me as I continue my scientific career. I also want to thank the members of the Weitzman lab, past and present, for their friendship and support - I would not have made it so far without you! I want to especially thank Daphne, who has been a wonderful mentor, helping me understand all the intricacies necessary for being a great scientist, especially writing and designing rigorous experiments. Daphne’s attention to detail nicely complemented Matt’s big picture view of my scientific development, supporting my development as a well- rounded scientist. I am also very grateful to Jen and Joe; whose help made the second project of this thesis possible. Additionally, I want to thank Alex and Matt C for many great and often crazy discussions inside the lab and out. They always reminded me what I love about science - that your craziest hypotheses are often the best, you just have to come up with a way to test them. Furthermore, I am thankful to our numerous collaborators, especially the members of the Garcia lab that have made my projects feasible with the magic that is mass spectrometry. I want to thank my committee members, Carolina Lopez, Sara Cherry, Paul Lieberman, and Paul Bates, for scientific input on my projects and their help in my career development. I am also exceedingly grateful to the amazing communities in CAMB, Microbiology, Protective Immunity, and Cancer Pathobiology for the exposure to great science and insightful discussions. A special thank-you goes to the CAMB office, Anna Kline, Meagan Schofer, Kathy O’Connor-Cooley, and Christina Strathearn, who have always been exceedingly helpful in anything grad school related, particularly in navigating Penn bureaucracy. iv I want to thank my previous mentors for helping me on my way to grad school. This is especially true for Walther Mothes, my advisor for my master’s thesis. Without his encouragement I would never have gone to Penn. Thank you for guiding me in the right direction! I also want to thank my friends back home and all the amazing people I have met here during my time at Penn. You have made grad school am amazing experience! I have met too many amazing people to possible thank them all - but I want to give a special shout out to Annie, Priya, Chris, Steve, the other Steve, Devin, Terra, Colleen, David, Somdutta, Ian, Kayla, and Vineet! Most importantly, I would like to thank my parents and my family. Without them this thesis, and the work it represents, would not have been possible. I know it has not always been easy for them that I chose to follow my dream on the other side of the world, but I would like to let them know how grateful I am for their unwavering support. I am also extremely grateful to my boyfriend Bobby for his love and support - I look forward to starting our next big adventure together! I also want to thank Bobby’s family for welcoming me into their midst. Finally, I want to thank my cat Cooper who has always been able to distract me when I most needed it. While he might not be entirely sane himself, he has definitely saved what is left of my sanity. v ABSTRACT ADENOVIRUS STRATEGIES FOR ALTERING THE CELLULAR ENVIRONMENT IN FAVOR OF INFECTION Christin Herrmann Matthew D. Weitzman, Ph.D. Viruses, as obligate intracellular pathogens, rely on their host cell for successful replication. Viruses have evolved different strategies to hijack and redirect cellular processes to benefit infection and overcome host immune responses. Understanding the mechanisms by which viruses exploit their host cells will reveal new targets for antiviral therapies. In addition, these studies can provide insights into the regulation of fundamental cellular processes. While much progress has been made in this area, many unexpected nuances of virus-host interaction are still being discovered. Here, we employed several strategies
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