Title Page
Contents
CHAPTER I. FOXO3a upregulates Nonhomologous End Joining (NHEJ), Nucleotide Excision Repair (NER), Mismatch Repair (MMR), and Base Excision Repair (BER) activities via transcriptional activation of XRCC4, XPC, MSH2 and APE1 11
ABSTRACT 12
INTRODUCTION 14
MATERIAL AND METHODS 17
RESULTS 28
DISCUSSION 50
REFERENCES 53
ABSTRACT IN KOREAN 59
CHAPTER II. FOXO3a Mediates Homologous Recombination Repair (HRR) via Transcriptional Activation of MRE11, BRCA1,BRIP1, and RAD50 61
ABSTRACT 62
INTRODUCTION 64
MATERIALS AND METHODS 67
RESULT 76
DISCUSSION 87
REFERENCES 91
ABSTRACT IN KOREAN 98
CHAPTER I 8
Figure 1. FOXO3a-dependent upregulation of the XRCC4, XPC, MSH2, and APE1 mRNAs in the tet-on flag h-FOXO3a transgenic mice. The tet-on flag-h-FOXO3a... 29
Figure 2. FOXO3a-dependent upregulation of the XRCC4, XPC, MSH2, and APE1 proteins, and NHEJ, NER, and BER activities in the tet-on flag-h-FOXO3a... 31
Figure 3. FOXO3a-dependent upregulation of NHEJ, NER, BER and MMR activities in the tet-on flag-h-FOXO3a transgenic mice. The tet-on flag-h-FOXO3a... 33
Figure 4. XRCC4 promoter activity was upregulated by FOXO3a binding to FOXO consensus sites of the promoter in MEFs of the tet-on flag-h-FOXO3a transgenic... 39
Figure 5. XPC promoter activity was upregulated by FOXO3a binding to FOXO consensus sites of the promoter in MEFs of the tet-on flag-h-FOXO3a transgenic... 40
Figure 6. MSH2 promoter activity was upregulated by FOXO3a binding to FOXO consensus sites of the promoter in MEFs of tet-on flag-h-FOXO3a transgenic... 42
Figure 7. APE1 promoter activity was upregulated by FOXO3a binding to FOXO consensus sites of the promoter in MEFs of the tet-on flag-h-FOXO3a transgenic... 43
Figure 8. Preparation of a tet-on flag-h-FOXO3a transgenic mice and a schematic diagram showing the relationship between FOXO3a and DNA repair.... 44
Figure 9. The tet-on flag-h-FOXO3a transgenic mice were used to test protein level of flag-h-FOXO3a in tissue obtained from brain, heart, thymus, pancreas,... 46
Figure 10. The protein level of Flag-h-FOXO3a, XRCC4. XPC, MSH2, and APE1 increased in a Doxycycline dose dependent manner in irradiated MEF cells and... 48
CHAPTER II 10
Figure 1. (A)The protein level of MRE11, BRCA1, BRIP1, RAD50, RAD51, FOXO3a, NBS1 BARD1, PALB2 in FOXO3a over-expressed or FOXO3a knock-down HDF... 77
Figure 2. The protein levels of MRE11, BRCA1, BRIP1, RAD50, RAD51, BRCA2, FOXO3a, and β-actin were measured by Western blotting. Cell extracts were... 79
Figure 3. MRE11, BRCA1, BRIP1, and RAD50 promoter activities were measured in young HDF cells transfected with increasing levels of wt-h-FOXO3a plasmid.... 81
Figure 4. FOXO3a-dependent upregulation of MRE11, BRCA1, BRIP1, and RAD50 mRNA and protein expression in tet-on flah-h-FOXO3a transgenic mice. The tet-... 84
Figure 5. In vitro HRR activity in DR-GFP and I-SceI plasmid transfected MEF and wt-MEF with DR-GFP (Addgene, #26475) plasmid and I-SceI (Addgene, #26477) plasmid.... 86
Figure 6. A schematic diagram showing up-regulation of HRR activity via transcriptional activation of target HRR genes by FOXO3a. 89