Molecular Evolution and Phylogenetic Importance of a Gamete Recognition Gene Zan Reveals a Unique Contribution to Mammalian Speciation
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Molecular evolution and phylogenetic importance of a gamete recognition gene Zan reveals a unique contribution to mammalian speciation. by Emma K. Roberts A Dissertation In Biological Sciences Submitted to the Graduate Faculty of Texas Tech University in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY Approved Robert D. Bradley Chair of Committee Daniel M. Hardy Llewellyn D. Densmore Caleb D. Phillips David A. Ray Mark Sheridan Dean of the Graduate School May, 2020 Copyright 2020, Emma K. Roberts Texas Tech University, Emma K. Roberts, May 2020 ACKNOWLEDGMENTS I would like to thank numerous people for support, both personally and professionally, throughout the course of my degree. First, I thank Dr. Robert D. Bradley for his mentorship, knowledge, and guidance throughout my tenure in in PhD program. His ‘open door policy’ helped me flourish and grow as a scientist. In addition, I thank Dr. Daniel M. Hardy for providing continued support, knowledge, and exciting collaborative efforts. I would also like to thank the remaining members of my advisory committee, Drs. Llewellyn D. Densmore III, Caleb D. Phillips, and David A. Ray for their patience, guidance, and support. The above advisors each helped mold me into a biologist and I am incredibly gracious for this gift. Additionally, I would like to thank numerous mentors, friends and colleagues for their advice, discussions, experience, and friendship. For these reasons, among others, I thank Dr. Faisal Ali Anwarali Khan, Dr. Sergio Balaguera-Reina, Dr. Ashish Bashyal, Joanna Bateman, Karishma Bisht, Kayla Bounds, Sarah Candler, Dr. Juan P. Carrera-Estupiñán, Dr. Megan Keith, Christopher Dunn, Moamen Elmassry, Dr. Adam Ferguson, Dr. Nicole Foley, James Francis, Dr. Sarah Fumagalli, Dr. Ken Griffith, Brandon Gross, Michaela Halsey, Dr. John Hanson, Dr. Lucas Heintzman, Dr. Tyla S. Holsomback, Dr. Howard M. Huynh, Dr. Narayan Kandel, Jennifer Korstian, Macy Krishnamoorthy, Dr. Neha Kumari, Mark Lee, Taylor Lenzmeier, Dr. Laramie Lindsey, Dr. Rita Magalhães, Sarah Mangum, Preston McDonald, Dr. Molly M. McDonough, Archana Muthu, Anisha Navlekar, Benneth Obitte, Dr. Nicté Ordóñez-Garza, Austin Osmanski, Dr. Julie Parlos, Nicole Paulat, Doug Perez, Dr. Kendra Phelps, Dr. R. Neal Platt, Andrea Reinhardt, Andres Rodriguez-Codero, Dr. Stephen A. Roussos, Megan Rowe, Oscar Sandate, Taylor Soniat, Dr. Cibele Sotero- Caio, Dr. Amanda Starr, Dr. Scott Starr, Dr. Jenny Strovas, Kevin Sullivan, Tim Sweeney, Iroro Tanshi, Dr. Courtney A. Thomason, Dr. Cody W. Thompson, Craig Tipton, Dr. Miryam Venegas-Anaya, Dr. Polrit Viravathana, Sarah Vrla, Dr. Lizz Waring, Rachael Weidmeier, and Emily Wright. I look forward to a lifetime of friendship with you all. ii Texas Tech University, Emma K. Roberts, May 2020 Thanks to Heath Garner, Kathy MacDonald, and the Natural Science Research Laboratory of the Museum of Texas Tech University, for assistance with tissue loans. Thank you to numerous co-authors who helped in data procurement and analysis as well as manuscript preparation for the research reported herein. I thank the TTU High Performance Computing Center for assistance with bioinformatics analyses. Thank you to the following professional and student organizations from which research funding was procured: National Institutes of Health, National Science Foundation, American Society of Mammalogists, Southwestern Association of Naturalists, Association of Biologists at Texas Tech University, Texas Tech University Graduate School Dissertation Completion Fellowship, Museum at Texas Tech University as well as the Department of Biological Sciences at Texas Tech University. Additionally, thanks to the Department of Biological Sciences and Graduate School of Texas Tech University for travel funding to present the research reported herein at scientific meetings. Additionally, thanks to the countless educators who with much patience taught me not only subject matter, but more importantly prepared me for both academia and life. Your words of encouragement, guidance, and knowledge did not go unheeded. With these sentiments I thank, among others, Carol Baker, Bob DeFazio, Dr. Ximena Bernal, Dr. Michael Dini, Dr. Lewis Held, Dr. Matt Johnson, David Schoen, Dr. Dylan Schwilk, and my parents (Terrence and Brenda Abrams). Finally, I would like to thank my family and close friends for their continued encouragement, reassurance, support, and love in all of my personal and professional endeavors, as well as the innumerable opportunities they provided for education and growth. Without all of them by my side, I would not have been successful in attaining a PhD, the greatest achievement of my academic career thus far. iii Texas Tech University, Emma K. Roberts, May 2020 TABLE OF CONTENTS ACKNOWLEDGEMENTS.........................................................................................ii LIST OF TABLES.......................................................................................................vi LIST OF FIGURES....................................................................................................vii I. INTRODUCTION.....................................................................................................1 Background Information....................................................................................1 Organization of Chapters.................................................................................10 II. RAPID DIVERGENCE OF A GAMETE RECOGNITION GENE PROMOTED MACROEVOLUTION OF EUTHERIA.........................................12 Abstract............................................................................................................12 Introduction......................................................................................................13 Materials and Methods.....................................................................................14 Experimental models/subjects..............................................................14 Database mining for Zan sequences.....................................................15 Sequence alignments............................................................................16 Phylogenetic analysis of Zan................................................................17 Phylogenetic comparisons....................................................................17 Selection tests.......................................................................................18 Characterization of zonadhesin D3 and D3p polypeptides...................19 Divergence rate comparisons, quantification, and statistical analyses................................................................................................19 Results...............................................................................................................20 Zan ontogeny.........................................................................................20 Zan phylogeny.......................................................................................23 Zan divergence......................................................................................26 Discussion.........................................................................................................28 III. GAMETE RECOGNITION GENE TREE YIELDS A ROBUST EUTHERIAN PHYLOGENY ACROSS TAXONOMIC LEVELS.......................63 Abstract.............................................................................................................63 Introduction.......................................................................................................64 Methods.............................................................................................................68 Results...............................................................................................................69 Magnordinal and Superordinal-level comparisons...............................69 Ordinal-level comparisons....................................................................70 Intra-ordinal comparisons.....................................................................72 iv Texas Tech University, Emma K. Roberts, May 2020 Discussion.........................................................................................................80 IV. ZAN VWD DOMAIN DUPLICATION AND DIVERGENCE REFLECT UNIQUE EVOLUTION OF A SPECIATION GENE IN MYOMORPH RODENTS...................................................................................................................96 Abstract.............................................................................................................96 Introduction......................................................................................................97 Methods...........................................................................................................101 Retrieval and comparison of Zan sequences.......................................101 Comparison of Zan in monotremes and placentals.............................102 Sequence alignments and phylogenetic analysis of Zan.....................102 Selection tests......................................................................................103 Results.............................................................................................................104 Retrieval and comparison of Zan sequences.......................................104 Comparisons of ZanL and Zan domain architecture...........................105 Phylogenetic and comparative