University of South Florida Scholar Commons Graduate Theses and Dissertations Graduate School January 2018 Characterization of the Transcriptional Elongation Factor ELL3 in B cells and Its Role in B-cell Lymphoma Proliferation and Survival Lou-Ella M.m. Alexander University of South Florida, [email protected] Follow this and additional works at: http://scholarcommons.usf.edu/etd Part of the Biology Commons Scholar Commons Citation Alexander, Lou-Ella M.m., "Characterization of the Transcriptional Elongation Factor ELL3 in B cells and Its Role in B-cell Lymphoma Proliferation and Survival" (2018). Graduate Theses and Dissertations. http://scholarcommons.usf.edu/etd/7119 This Dissertation is brought to you for free and open access by the Graduate School at Scholar Commons. It has been accepted for inclusion in Graduate Theses and Dissertations by an authorized administrator of Scholar Commons. For more information, please contact [email protected]. Characterization of the Transcriptional Elongation Factor ELL3 in B cells and Its Role in B-cell Lymphoma Proliferation and Survival by Lou-Ella M. M. Alexander A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy Department of Cell Biology, Microbiology and Molecular Biology College of Arts and Sciences University of South Florida Major Professor: Kenneth L. Wright, Ph.D. Sheng Wei, M.D. Shari A. Pilon-Thomas, Ph.D. Mark G. Alexandrow, Ph.D. Date of Approval: November 29, 2017 Keywords: Germinal center, Eleven-nineteen Lysine-rich Leukemia, cell cycle Copyright © 2017, Lou-Ella M. M. Alexander DEDICATION This dissertation is dedicated to my mother, Jessica Alexander-Ogenia, and my father, Otmar Alexander, who provided me with the opportunity to get a great education abroad and taught me the importance of hard work, determination, patience and sacrifice to achieve one’s dreams. Mama i Tata, ma bai p’é i porfin ma logré! ACKNOWLEDGMENTS I would like to take the opportunity to thank my major professor Dr. Kenneth Wright, for his endless guidance and contributions towards my development into an independent cancer researcher. I am grateful to the University of South Florida and the Cancer Biology Ph.D. program for the opportunity to join the graduate program and participate in cutting edge cancer research. I would also like to thank my committee members Dr. Sheng Wei, Dr. Shari Pilon- Thomas and Dr. Mark Alexandrow, for their critical scientific input and guidance. I am also grateful to Barbara Vilen, Ph.D. for agreeing to serve as my external chair. I have to thank past and present members of the Wright laboratory, colleagues and friends for their scientific contributions, constructive criticism and encouragement throughout this process. Special thanks to my friends, Rydienne Cornelia, Jeanise Job, Victoria Bryant, Gabriela Wright and January Watters for their support during both my undergraduate and graduate careers. Being away from my family, it has been amazing to have you all by my side. Thank you for all the wise words, the listening ear and most importantly your friendship throughout the years. I am eternally grateful to the Ogenia and Alexander family in Curaçao, Aruba and the Netherlands who have made my virtual attendance to family gatherings possible and send care packages. Your efforts have brought me lots of comfort when I was missing home and always brightened my day. I love you all immensely! I am extremely grateful to my brother Alexis Alexander, my sister-in-law Daniece Alexander-Reed and my niece Xaenah Alexander for showing their support through phone calls, pictures and videos. Unknowingly, every single one came at a time that I need it most and provided some much needed comedic relief. I love you all very much! Words cannot describe my gratitude to the love of my life, Craig George for his continued and unfailing love, support and understanding during the completion of this Ph.D. degree. You were always there to comfort me and help keep things in perspective. Mi stimabu un mundu and I cannot wait to see what the future has in store for us. An immense thank you to the George and Pfahler families, you have all been a source of support and encouragement as I completed my studies and transitioned to Atlanta. You have all opened your hearts and welcomed me into the family with open arms. I am extremely grateful and blessed to have you all in my life! I am forever grateful to my parents Jessica Alexander-Ogenia and Otmar Alexander who have been an immense force behind me throughout my whole career. Their love, support, patience and sacrifices are unmatchable and I do not have the words to thank you enough. I love you both from the bottom of my heart. Masha masha danki Mama I Tata! TABLE OF CONTENTS LIST OF TABLES ......................................................................................................................... iv LIST OF FIGURES .........................................................................................................................v LIST OF ABBREVIATIONS ....................................................................................................... vii ABSTRACT ................................................................................................................................... xi CHAPTER I: INTRODUCTION .....................................................................................................1 1.1 Hematopoiesis ................................................................................................................1 1.2 B cell Development........................................................................................................3 1.2.1 Antigen-Independent Phase ............................................................................4 1.2.1.1 HSC to MPP Transition ...................................................................4 1.2.1.2 MPP to ELP/CLP Differentiation ....................................................6 1.2.1.3 CLP to Mature B cell Differentiation ..............................................7 1.2.2 Antigen-Dependent Phase .............................................................................11 1.2.2.1 T-cell Independent Phase ...............................................................11 1.2.2.2 T-cell Dependent Phase .................................................................12 1.2.3 Antibody Diversity........................................................................................13 1.2.4 Transcriptional Regulators in T-cell Dependent Phase ................................14 1.3 Lymphomas..................................................................................................................19 1.3.1 Lymphoma Statistics .....................................................................................20 1.3.2 B cell Lymphomas ........................................................................................21 1.3.2.1 Cause of B cell Lymphomas ..........................................................21 1.3.2.2 B cell Lymphoma Classification ....................................................22 1.4 Transcription ................................................................................................................27 1.5 RNA Polymerase II Transcription ...............................................................................27 1.5.1 Transcriptional Initiation ..............................................................................28 1.5.2 Transcriptional Elongation............................................................................29 1.5.2.1 Promoter Clearance ........................................................................30 1.5.2.2 Promoter Proximal Pausing ...........................................................30 1.5.2.3 Effective Elongation ......................................................................31 1.5.3 Transcriptional Termination .........................................................................32 1.6 ELL Family Members ..................................................................................................33 1.6.1 ELL ...............................................................................................................33 1.6.1.1 ELL Structure.................................................................................33 1.6.1.2 ELL Functions ...............................................................................33 i 1.6.2 ELL2 .............................................................................................................34 1.6.2.1 ELL2 Structure...............................................................................34 1.6.2.2 Cell-type Specific Function of ELL2 .............................................34 1.6.3 ELL3 .............................................................................................................35 1.6.3.1 ELL3 Structure...............................................................................35 1.6.3.2 Cell-type Specific Functions of ELL3 ...........................................35 1.6.4 SEC-like Complexes .....................................................................................35 CHAPTER II: MATERIALS & METHODS ................................................................................37 2.1 Cell Lines and Reagents..............................................................................................37 2.2 Peripheral Blood Mononuclear Cell Isolation ............................................................37