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UC San Diego UC San Diego Electronic Theses and Dissertations Title Ionizing radiation induces ATM-independent degradation of p21Cip1 in transformed cells Permalink https://escholarship.org/uc/item/7m49k70z Author Stuart, Scott Publication Date 2008 Peer reviewed|Thesis/dissertation eScholarship.org Powered by the California Digital Library University of California UNIVERSITY OF CALIFORNIA, SAN DIEGO Ionizing Radiation Induces ATM-independent Degradation of p21Cip1 in Transformed Cells A dissertation submitted in partial satisfaction of the requirements for the degree Doctor of Philosophy in Biomedical Sciences by Scott Stuart Committee in charge: Professor Jean J.Y. Wang, Chair Professor Steven F. Dowdy Professor Geoff Wahl Professor Tony Wynshaw-Boris Professor Huilin Zhou 2008 © Scott A. Stuart, 2008 All rights reserved. The Dissertation of Scott Andrew Stuart is approved, and it is acceptable in quality and form for publication on microfilm and electronically: Chair University of California, San Diego 2008 iii Dedication This dissertation is dedicated in memory of Evelyn F. Mahle and Natalie J. Stuart. iv Table of Contents Signature Page ......................................................................................................... iii Dedication ............................................................................................................... iv Table of Contents .................................................................................................... v List of Abbreviations .............................................................................................. vii List of Figures ......................................................................................................... x Acknowledgements ................................................................................................. xiii Vita .......................................................................................................................... xiv Abstract of the Dissertation ..................................................................................... xvi Chapter 1: Introduction ........................................................................................ 1 References .................................................................................................... 11 Chapter 2: Ionizing Radiation Induces ATM-independent Degradation of p21cip1 in Transformed Cells ............................................................................... 16 Abstract .......................................................................................................... 16 Introduction .................................................................................................... 17 Experimental Procedures ............................................................................... 18 Results ............................................................................................................ 24 Discussion ...................................................................................................... 33 References ...................................................................................................... 45 Chapter 3: Transformation of E2A-Deficient Pluripotent Progenitors by BCR-ABL Generates Leukemic Stem Cell in the GMP Compartment ............. 49 Abstract .......................................................................................................... 49 v Introduction .................................................................................................... 50 Experimental Procedures ............................................................................... 52 Results ............................................................................................................ 57 Discussion ...................................................................................................... 65 References ...................................................................................................... 90 Chapter 4: Discussion .............................................................................................. 94 References ..................................................................................................... 105 vi List of Abbreviations 4-OHT - 4-hydroxytamoxifen A-T - Ataxia-Telangiectasia Abl - Abelson murine leukemia viral oncogene homolog 1 ATM - Ataxia-Telangiectasia Mutated ATP - Adenine Triphosphate ATR - ATM/Rad3 Related BCR-ABL - Breakpoint Cluster Region-Abelson fusion protein BM - Bone Marrow BrdU - Bromodeoxyuridine CD - Cluster of Differentiation cdc - cell division cycle cdk - cyclin-dependent kinase Chk1 - Checkpoint kinase 1 Chk2 - Checkpoint kinase 2 CHX - Cyclohexamide Cip - Cdk-interacting protein CKI - Cdk Inhibitor protein CML - Chronic Myelogenous Leukemia CSN - COP9 signalosome Cul - Cullin - vii DCAF - DDB1-Cul4 associated factors DDB1 - DNA damage binding protein 1 DNA - Deoxyribonucleic acid DNA-PK - DNA-dependent protein kinase E1 - E1 Ubiquitin activating enzyme E2 - E2 Ubiquitin conjugating enzyme E3 - E3 Ubiquitin ligase ER - Estrogen Receptor FACS - Fluorescence Activated Cell Sorting GFP - Green Fluorescence Protein GMP - Granulocyte Macrophage Progenitor Gy - Gray HA - Hemagglutinin HEK - Human Embryonic Kidney HFF - Human Foreskin Fibroblast INK4 - Inhibitors of cdk4 IR - Ionizing Radiation IRES - Internal Ribosome Entry Site LEF - Lymphoid Enhancer Factor MEF - Mouse Embryonic Fibroblast MEP - Myeloid Erythroid Progenitor mHPC - murine hematopoietic progenitor cell viii MSCV - Murine Stem Cell Virus Nbs1 - Nijmegen breakage syndrome 1 p210 - BCR-ABL 210 kilodalton protein PB - Peripheral blood PCNA - Proliferating Cell Nuclear Antigen PIKK - PI3-like protein kinase Rb - Retinoblastoma protein SCF - Skp1/Cul1/Rbx1/F-box complex shRNA - short hairpin RNA siRNA - small interfering RNA Skp2 - S-phase kinase associated protein 2 Smc1 - Structural maintenance of chromosomes protein 1 SV40 - Simian Virus 40 TCF - Transcription Factor Gene UV - Ultraviolet radiation WBC - White blood cell ZVAD-fmk - N-benzyloxycarbonyl-valyl-alanyl-aspartyl-fluoromethylketone ix
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