Monitoring homology search during DNA double-strand break repair in vivo:
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-169454 Inhaltsverzeichnis |
| Physical Description: | 108 S. graph. Darst. |
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Record in the Search Index
| _version_ | 1804152199685603328 |
|---|---|
| adam_text | MONITORING HOMOLOGY SEARCH
DURING DNA DOUBLE-STRAND BREAK REPAIR IN VIVO
DISSERTATION DER FAKULTAT FOR BIOLOGIE
DER LUDWIG-MAXIMILIANS-UNIVERSITAT MUNCHEN
VORGELEGT VON
JORG RENKAWITZ, M.SC. BIOCHEMIE
JUNI 2013
HTTP://D-NB.INFO/1051818214
TABLE OF CONTENTS
TABLE OF CONTENTS
1 SUMMARY 3
2 INTRODUCTION 4
2.1 HOMOLOGOUS RECOMBINATION 5
2.2
SACCHAROMYCES CEREVISIAE
REPRESENTS A MODEL ORGANISM
TO STUDY
HOMOLOGOUS RECOMBINATION 7
2.3 THE RECOMBINASE RAD51 10
2.4 REGULATION OF HOMOLOGOUS RECOMBINATION 12
2.5 HOMOLOGY SEARCH 15
3 AIMS OF THIS STUDY 19
4 RESULTS 20
4.1 HOMOLOGY SEARCH CAN BE MONITORED
IN
VIVO
BY RAD51 CHIP 20
4.1.1 BROAD DISTRIBUTION OF RAD51 CHIP SIGNALS UPON A SINGLE DSB 20
4.1.2 DSB-DISTANT RAD51 CHIP SIGNALS ARE SPECIFIC 23
4.1.3 DSB-DISTANT RAD51 CHIP SIGNALS DEPEND ON RAD52 AND RAD55/RAD57..
25
4.1.4 DSB-DISTANT RAD51 CHIP SIGNALS ARE A GENERAL PHENOMENON UPON
SINGLE DSBS IN SACCHAROMYCES CEREVISIAE 26
4.1.5 DSB-DISTANT RAD51 CHIP SIGNALS EXCEED THE AREA OF RESECTION 28
4.1.6 DSB-DISTANT RAD51 CHIP SIGNALS ARE INDICATIVE OF RECOMBINATION
COMPETENCE 32
4.1.7 DSB-DISTANT RAD51 CHIP SIGNALS ARE TRACEABLE DURING ACTIVE
REPAIR..36
4.1.8 DSB-DISTANT RAD51 CHIP SIGNALS DEPEND ON A DNA-BINDING SITE OF
RAD51 39
4.2 NUCLEAR ORGANIZATION INFLUENCES HOMOLOGY SEARCH 41
I 4.2.1 THE RE ELEMENT CAN GUIDE HOMOLOGY SEARCH IN TRANS 41
4.2.2 CENTROMERES CAN GUIDE HOMOLOGY SEARCH IN TRANS 45
4.2.3 HOMOLOGY SEARCH UPON SINGLE DSBS IN PROXIMITY OF DIFFERENT NUCLEAR
LANDMARKS 49
4.3 COMPONENTS AND REQUIREMENTS OF
HOMOLOGY SEARCH 53
4.3.1 ROLE OF THE SINGLE-STRAND BINDING PROTEIN RPA DURING HOMOLOGY
SEARCH.
53
4.3.2 EFFICIENT HOMOLOGY SEARCH REQUIRES RAD54 56
4.3.3 PHOSPHORYLATION OF HISTONE 2A FOLLOWS HOMOLOGY SEARCH 58
5 DISCUSSION 64
5.1 MONITORING HOMOLOGY SEARCH
IN VIVO
64
5.2 CONFLICTING MODELS FOR HOMOLOGY SEARCH 66
5.3 HOMOLOGY SEARCH IS INFLUENCED BY THE NUCLEAR
ORGANIZATION 68
5.4 R
EVISITING EARLIER
D
ATA IN
V
IEW OF
H
OMOLOGY
S
EARCH BEING
I
NFLUENCED BY
N
UCLEAR
O
RGANIZATION 72
5.5 FACILITATED 3D/1D HOMOLOGY SEARCH
MODEL 74
6 MATERIALS AND METHODS 78
6.1 MICROBIOLOGY 78
6.1.1 ESCHERICHIA COLI TECHNIQUES 78
6.1.2 SACCHAROMYCES CEREVISIAE TECHNIQUES 79
6.2 MOLECULAR BIOLOGICAL TECHNIQUES 84
6.2.1 POLYMERASE CHAIN REACTION (PCR) 84
6.2.2 MOLECULAR CLONING 86
1
TABLE OF CONTENTS
6.3 BIOCHEMISTRY AND CELL BIOLOGY TECHNIQUES 87
6.3.1 PROTEIN METHODS 87
6.3.2 INDUCTION OF SINGLE DSBS IN VIVO 89
6.3.3 CHROMATIN-LMMUNOPRECIPITATIONS (CHIP) 89
6.3.4 CHIP-ON-CHIP ANALYSIS 92
6.4 COMPUTER-AIDED ANALYSIS 93
6.5
S
TATISTICAL
A
NALYSIS 93
7 REFERENCES 94
8 ABBREVIATIONS 103
9 FIGURE INDEX 104
10 ACKNOWLEDGEMENTS 106
11 CURRICULUM VITAE 107
2
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| spelling | Renkawitz, Jörg Verfasser aut Monitoring homology search during DNA double-strand break repair in vivo von Jörg Renkawitz 2013 108 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-169454 http://edoc.ub.uni-muenchen.de/16945/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-169454 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027296939&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Renkawitz, Jörg Monitoring homology search during DNA double-strand break repair in vivo |
| subject_GND | (DE-588)4113937-9 |
| title | Monitoring homology search during DNA double-strand break repair in vivo |
| title_auth | Monitoring homology search during DNA double-strand break repair in vivo |
| title_exact_search | Monitoring homology search during DNA double-strand break repair in vivo |
| title_full | Monitoring homology search during DNA double-strand break repair in vivo von Jörg Renkawitz |
| title_fullStr | Monitoring homology search during DNA double-strand break repair in vivo von Jörg Renkawitz |
| title_full_unstemmed | Monitoring homology search during DNA double-strand break repair in vivo von Jörg Renkawitz |
| title_short | Monitoring homology search during DNA double-strand break repair in vivo |
| title_sort | monitoring homology search during dna double strand break repair in vivo |
| topic_facet | Hochschulschrift |
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