Dissection of the topology, structure and function of the INO80 chromatin remodeler:
Gespeichert in:
| Beteilige Person: | |
|---|---|
| Format: | Hochschulschrift/Dissertation Buch |
| Sprache: | Englisch |
| Veröffentlicht: |
2013
|
| Schlagwörter: | |
| Links: | https://nbn-resolving.org/urn:nbn:de:bvb:19-164074 http://d-nb.info/1046503081/34 http://edoc.ub.uni-muenchen.de/16407/ http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027097222&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| Umfang: | 113 Blatt Ill., graph. Darst. |
| Format: | Langzeitarchivierung gewährleistet, LZA |
Internformat
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| 100 | 1 | |a Tosi, Alessandro |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Dissection of the topology, structure and function of the INO80 chromatin remodeler |c Alessandro Tosi |
| 264 | 1 | |c 2013 | |
| 300 | |a 113 Blatt |b Ill., graph. Darst. | ||
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Datensatz im Suchindex
| DE-BY-TUM_katkey | 1993115 |
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| adam_text | TABLE OF CONTENTS 4
1 TABLE OF CONTENTS
1 TABLE OF CONTENTS 4
2 SUMMARY 7
3 INTRODUCTION ; 9
3.1 DYNAMIC CHROMATIN
ENVIRONMENT 9
3.2 SWI2/SNF2 REMODELERS 11
3.3 CHROMATIN REMODELERS 13
3.4 THE IN080/SWR1 FAMILY 14
3.5 IN080 COMPLEX 16
3.5.1 THE COMPONENTS OF
THE IN080 COMPLEX 16
3.5.2 THE CHROMATIN REMODELING COMPLEX
IN080 IS INVOLVED IN DNA
PROCESSING AND METABOLISM
20
3.5.3 IN080 MEDIATES CHECKPOINT PATHWAYS 21
3.6 HYBRID APPROACHES HELP
TO DISSECT THE
MOLECULAR ARCHITECTURE OF LARGE
COMPLEXES 22
4 RESULTS 25
4.1 RECONSTITUTE OF A NUCLEOSOME 25
4.2 A NOVEL
PURIFICATION PROCEDURE OF IN080
IMPROVES COMPLEX HOMOGENEITY 27
4.3 NANOBODIES AGAINST THE
IN080 COMPLEX 31
4.4 ASSESSMENT OF THE
ACTIVITY OF THE PURIFIED
IN080 33
4.5 CHEMICAL CROSS-UNKING AND
MASS SPECTROMETRY ANALYSIS OF
THE IN080
COMPLEX 35
4.5.1 MAPPING OF SUBUNIT INTERACTIONS BY CROSS-UNKING AND MASS
SPECTROMETRY 35
4.5.2 SUBUNIT TOPOLOGY AND STRUCTURAL MODULES
OF IN080 38
4.6 VALIDATION OF I N080
S
MODULES
IN VIVO
40
4.7 STRUCTURE OF THE
IN080 COMPLEX 41
4.7.1 ELECTRON MICROSCOPY OF IN080 41
4.7.2 TOWARDS A CRYSTAL STRUCTURE OF IN080 43
4.8 RVB1/2 FORM
A HETERO-DODECAMER 43
4.8.1 RVB1/2 IS COMPOSED OF
TWO HEXAMERIC RINGS
IN IN080 43
4.8.2 RVB1/2 ASSEMBLE AS HETERO-HEXAMERS
INTERACTING VIA THE
DOMAIN 2 WITHIN IN080 44
HTTP://D-NB.INFO/1048302083
TABLE OF CONTENTS
5
4.9 THE CATALYTIC
CORE OF IN080 47
4.9.1 THE SWI2/SNF2 DOMAIN
OF IN080 47
4.9.2 EXPRESSION AND PURIFICATION STUDIES
OF THE ATPASE
DOMAIN OF INO80
WITH IES2 AND RVBI 48
4.10 LOCALIZATION OF THE ARP8-,ARP5- AND NHPIO-MODULE 48
4.11 THE NHPIO-MODULE 50
4.11.1 RECONSTITUTION OF THE NHPIO
MODULE: NHP10-IES1-IES3-IES5-INO80
(N-TERMINUS) 50
4.11.2 THE NHPIO SUB-COMPLEX FORMS
A STABLE DNA COMPLEX 53
4.12 DISSECTING THE
FUNCTION AND ACTIVITY
OF IN080-MODULES 55
4.12.1 FUNCTIONAL CHARACTERIZATION OF IN080-MODULES 55
4.12.2 INTERACTION AND VISUALIZATION OF AN IN080-NUCLESOME
COMPLEX 57
4.13 IN080 FORMS A STABLE
COMPLEX WITH THE
MECI COMPLEX 61
5 DISCUSSION 63
5.1 HYBRID VIEW ON IN080 63
5.2 THE CHROMATIN
REMODELER IN080 64
5.3 STRUCTURE OF
RVB1/2
IN THE IN080
COMPLEX 65
5.4 THE NHPIO-MODULE 68
5.5 THE ARP8-MODULE 69
5.6 THE NUCLEOSOME
REMODELER IN080 70
5.7 CHROMATIN REGULATORS
FACILITATE TRANSCRIPTION 75
6 MATERIAL AND METHODS 77
6.1 MATERIALS 77
6.2 YEAST STRAINS 78
6.3 PLASMID LISTS 78
6.4 OLIGONUCLEOTIDE UST 79
6.5 BUFFER UST 81
6.6 MOLECULAR BIOLOGY METHODS 82
6.6.1 CLONING IN BACTERIA 82
6.6.2 CLONING IN YEAST 83
6.7 PROTEIN EXPRESSION AND
PURIFICATION 83
6.7.1 PROTEIN EXPRESSION IN E.
COLI 83
TABLE OF CONTENTS 6
6.7.2 PROTEIN EXPRESSION IN INSECT CELLS 84
6.7.3 CULTURING OF YEAST 84
6.7.4 SDS-PAGE AND WESTERN BLOT ANALYSIS 85
6.7.5 SILVER STAINING 86
6.7.6 PURIFICATION OF HISTONES
AND RECONSTITUTION OF NUCLEOSOMES 87
6.7.7 ESTABLISHMENT OF AN IN080
PURIFICATION PROTOCOL 89
6.7.8 PURIFICATION OF THE SWI2/SNF2
SUB-COMPLEX 92
6.7.9 PURIFICATION OF THE NHPIOSUBCOMPLEX 92
6.8 GENERATION OF
IN080 BINDING NANOBODIES 93
6.9 CROSS-UNKING AND MASS
SPECTROMETRY 94
6.9.1 TITRATION OF CROSS-LINKER 94
6.9.2 SAMPLE PREPARATION FOR MASS SPECTROMETRY ANALYSIS 94
6.9.3 SAMPLE ANALYSIS 95
6.10 ANALYSIS OF
SUBUNIT COMPOSITION
OF INOSO S
KNOCK-OUT MUTANTS 96
6.11 ANALYSIS OF DISTANT RESTRAINTS 96
6.12 FUNCTIONAL ASSAYS 97
6.12.1 ELECTROPHORETIC MOBILITY SHIFT ASSAYS 97
6.12.2 REMODELING ASSAY 97
6.12.3 ATP HYDROLYSIS ASSAYS 98
7 REFERENCES 99
8 CURRICULUM VITAE 111
9 ACKNOWLEDGMENTS
113
|
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| indexdate | 2024-12-20T16:51:37Z |
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| language | English |
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| spellingShingle | Tosi, Alessandro Dissection of the topology, structure and function of the INO80 chromatin remodeler |
| subject_GND | (DE-588)4113937-9 |
| title | Dissection of the topology, structure and function of the INO80 chromatin remodeler |
| title_auth | Dissection of the topology, structure and function of the INO80 chromatin remodeler |
| title_exact_search | Dissection of the topology, structure and function of the INO80 chromatin remodeler |
| title_full | Dissection of the topology, structure and function of the INO80 chromatin remodeler Alessandro Tosi |
| title_fullStr | Dissection of the topology, structure and function of the INO80 chromatin remodeler Alessandro Tosi |
| title_full_unstemmed | Dissection of the topology, structure and function of the INO80 chromatin remodeler Alessandro Tosi |
| title_short | Dissection of the topology, structure and function of the INO80 chromatin remodeler |
| title_sort | dissection of the topology structure and function of the ino80 chromatin remodeler |
| topic_facet | Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:19-164074 http://d-nb.info/1046503081/34 http://edoc.ub.uni-muenchen.de/16407/ http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027097222&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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