Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae:
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
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Berlin
2015
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| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 111 Seiten Illustrationen, Diagramme |
Internformat
MARC
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| 245 | 1 | 0 | |a Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae |c von Tanja Vieregge |
| 264 | 1 | |a Berlin |c 2015 | |
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| 776 | 0 | 8 | |i Reproduziert als |a Vieregge, Tanja |t Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae |d Berlin : MIK-Center GmbH, 2015 |h 2 Mikrofiches |w (DE-604)BV043022571 |
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Datensatz im Suchindex
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| adam_text | TABLE OF CONTENTS
TABLE OF CONTENTS
ABSTRACT 2
ZUSAMMENFASSUNG 3
TABLE OF CONTENTS 5
LIST OF TABLES 8
LIST OF FIGURES 9
ABBREVIATIONS 11
1 INTRODUCTION 13
1.1 THE ORGANIZATION OF DNA IN EUKARYOTES 13
1.2 ATP-DEPENDENT REMODELING COMPLEXES 15
1.3 HISTONE MODIFICATIONS 17
1.3.1 MODIFICATIONS OF THE HISTONE CORE DOMAIN 18
1.3.2 HISTONE PHOSPHORYLATION 20
1.4 HISTONE VARIANTS 21
1.5 HISTONE VARIANT H2A.Z 23
1.5.1 THE H2A.Z CONTAINING NUCLEOSOME 24
1.5.2 GENOME-WIDE LOCALIZATION OF H2A.Z 26
1.5.3 POSTTRANSLATIONAL MODIFICATIONS ON H2A.Z 27
1.5.4 H2A.Z DYNAMICS: INCORPORATION AND EVICTION 28
1.5.5 H2A.Z FUNCTION 31
1.5.5.1 TRANSCRIPTION 31
1.5.5.2 HETEROCHROMATIN FORMATION AND BOUNDARY FUNCTION 33
1.5.5.3 GENOMIC STABILITY AND DNA REPAIR 34
1.6 OUTLINE OF THIS THESIS 34
2 MATERIAL AND
METHODS 36
2.1 MEDIA AND GROWTH CONDITIONS 36
2.2 ESCHERICHIA COLI STRAINS 36
2.3 SACCHAROMYCES CEREVISIAE STRAINS 36
2.4 GENERATION OF YEAST STRAINS 38
2.4.1 DNA TECHNIQUES IN S. CEREVISIAE 38
2.4.2 GENETIC CROSSES, SPORULATION AND TETRAD DISSECTION OF S.
CEREVISIAE 39
2.5 MOLECULAR CLONING 39
5
HTTP://D-NB.INFO/1075495628
TABLE OF CONTENTS
2.6 PROTEIN PURIFICATION ACCORDING TO WATERBORG 41
2.7 TANDEM AFFINITY PURIFICATION (TAP) 42
2.8 MASS SPECTROMETRY ANALYSIS 43
2.9 GENERATION AND PURIFICATION OF A-HTZL-T108PH ANTIBODIES 44
2.10 CHROMATIN IMMUNOPRECIPITATION 45
2.11 RNA PURIFICATION, CDNA PREPARATION AND QPCR 46
2.12 QUANTITATIVE REAL-TIME PCR 47
2.13 NEXT GENERATION SEQUENCING 48
2.14 PROTEIN EXTRACTION OF 5. CEREVISIAE 49
2.15 IMMUNOPRECIPITATION (IP) 49
2.16SDS-PAGE AND IMMUNOBLOTTING (WESTERN BLOT) 49
2.17 FACS (FLUORESCENCE ASSOCIATED CELL SORTING) 50
2.18 HISTONE REPLACEMENT ASSAY 50
2.19 CELL FRACTIONATION 51
3 RESULTS 53
3.1 IDENTIFICATION OF NOVEL PTMS ON HTZL 53
3.1.1 LARGE-SCALE PURIFICATION OF HTZL FROM YEAST 53
3.1.2 MASS SPECTROMIC ANALYSIS OF TAP-PURIFIED HTZL 55
3.2 VALIDATION OF HTZL T108PH USING A SPECIFIC ANTIBODY 57
3.2.1 GENERATION AND PURIFICATION OF THE T108PH-ANTIBODY 57
3.2.2 DETECTION OF THE HTZL-T108 PHOSPHORYLATION IN VIVO 59
3.3 HTZL-T108 MUTATIONS LEAD TO GLOBAL DEREALIZATION OF HTZL 59
3.3.1 MUTATION OF HTZL-T108 LEADS TO A DEREALIZATION OF HTZL FROM THE
TSS 60
3.3.2 HTZL-T108 MUTATIONS AFFECT HTZL LOCALIZATION AT CENTROMERES AND
TELOMERES 62
3.4 MUTATIONS OF HTZL-T108 DO NOT AFFECT THE OCCUPANCY OF HTZL IN
CHROMATIN 64
3.5 THE HTZL-T108E MUTANT REDUCED SWRL-MEDIATED INCORPORATION OF
HTZL/H2B
DIMERS 65
3.6 THE PHOSPHORYLATION OF HTZL-T108 IS LOCALIZED TO SPECIFIC GENOMIC
FEATURES 66
3.6.1 HTZL-T108 PHOSPHORYLATION IS ENRICHED AT PROMOTER REGIONS 67
3.6.2 HTZL-T108 PHOSPHORYLATION IS ENRICHED AT CENTROMERIC AND TELOMERIC
REGIONS 67
3.7 DYNAMICS OF HTZL-T108 PHOSPHORYLATION UNDER STRESS CONDITIONS 69
3.7.1 HTZL-T108E AND HTZL-T108A CAUSE CHANGES IN GENE INDUCTION OF
STRESS-INDUCIBLE
GENES 69
3.7.2 ANALYSIS OF HTZL-T108 PHOSPHORYLATION AT THE TSS OF STRESS-INDUCED
GENES 72
3.8 THE HTZL-T108E MUTANT CAUSES GENOMIC INSTABILITY IN COMBINATION WITH
H4
MUTATIONS 73
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TABLE OF CONTENTS
3.9 PHENOTYPIC CHARACTERIZATION OF HTZL-T108 MUTANTS 75
3.9.1 HTZL-T108 MUTATIONS ENHANCE THE TEMPERATURE SENSITIVITY OF CTKA
CELLS 76
3.9.2 MUTATIONS OF HTZL-T108 HAVE NO EFFECT WHEN COMBINED WITH RPD3A 77
3.9.3 MUTATIONS OF HTZL-T108 DO NOT ENHANCE THE HU SENSITIVITY OF ARP8A
CELLS 78
3.9.4 MUTATIONS OF HTZL-T108 DO NOT CAUSE TEMPERATURE SENSITIVITY 79
3.9.5 MUTATIONS OF HTZL-T108 DO NOT CAUSE SENSITIVITY TO GENOTOXIC AND
STRESS-PROVOKING
AGENTS 79
3.10 IDENTIFICATION OF THE HTZL-T108PH KINASE 80
3.10.1 PHOSPHORYLATION OF HTZL-T108 BY THE RNA POLYMERASE II KINASES
CTKL AND KIN28
81
3.10.2 HTZL-T108 IS NOT PHOSPHORYLATED BY THE STRESS-RELATED KINASES
SNFL, DBF2 AND
HOGL 82
4 DISCUSSION 84
4.1 POSTTRANSLATIONAL MODIFICATIONS ON HTZL 84
4.2 POTENTIAL STRUCTURAL CONSEQUENCES OF HTZL-T108 PHOSPHORYLATION
WITHIN THE
NUCLEOSOME 86
4.3 INTERACTION BETWEEN HTZL-T108PH AND SWR1 88
4.4 HOW DOES HTZL-T108 PHOSPHORYLATION AFFECT GENE EXPRESSION? ...89
4.5 TIMING OF HTZL-T108 PHOSPHORYLATION 90
4.6 PUTATIVE KINASE FOR HTZL-T108 PHOSPHORYLATION 92
4.7 SUMMARY OF THE MAIN RESULTS 93
5 REFERENCES 95
6 APPENDIX 108
7 ADDENDUM 109
EIDESSTATTLICHE ERKLARUNGEN ILL
7
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| spelling | Vieregge, Tanja Verfasser (DE-588)1069681512 aut Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae von Tanja Vieregge Berlin 2015 111 Seiten Illustrationen, Diagramme txt rdacontent n rdamedia nc rdacarrier Dissertation Humboldt-Universität zu Berlin 2015 (DE-588)4113937-9 Hochschulschrift gnd-content Reproduziert als Vieregge, Tanja Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae Berlin : MIK-Center GmbH, 2015 2 Mikrofiches (DE-604)BV043022571 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027943124&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Vieregge, Tanja Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae |
| subject_GND | (DE-588)4113937-9 |
| title | Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae |
| title_auth | Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae |
| title_exact_search | Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae |
| title_full | Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae von Tanja Vieregge |
| title_fullStr | Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae von Tanja Vieregge |
| title_full_unstemmed | Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae von Tanja Vieregge |
| title_short | Identification and characterization of posttranslational modifications on the histone variant H2A.Z in Saccharomyces cerevisiae |
| title_sort | identification and characterization of posttranslational modifications on the histone variant h2a z in saccharomyces cerevisiae |
| topic_facet | Hochschulschrift |
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