Pericentriolar material assembly in Caenorhabditis elegans:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2015
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | VIII, 93, 2 S. Ill., graph. Darst. CD-ROM (12 cm) |
Internformat
MARC
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| 245 | 1 | 0 | |a Pericentriolar material assembly in Caenorhabditis elegans |c von Oliver Wüseke |
| 264 | 1 | |c 2015 | |
| 300 | |a VIII, 93, 2 S. |b Ill., graph. Darst. |e CD-ROM (12 cm) | ||
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| 502 | |a Dresden, Techn. Univ., Diss., 2015 | ||
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-028117541 | ||
Datensatz im Suchindex
| _version_ | 1804174878894456832 |
|---|---|
| adam_text | CONTENTS
1 INTRODUCTION 1
1.1 THE CYTOSKELETON 1
1.2 THE MICROTUBULE NETWORK 2
1.2.1 MICROTUBULE-ASSOCIATED PROTEINS 2
1.2.2 MICROTUBULE-ORGANIZING CENTERS 3
1.3 THE CENTROSOME 4
1.3.1 CENTROSOME STRUCTURE 4
1.3.2 CENTROSOME DYNAMICS 7
1.4 C. ELEGANS AS A MODEL SYSTEM TO STUDY PCM ASSEMBLY 9
1.5 AIM OF THIS STUDY 10
2 RESULTS - CYTOPLASMIC INTERACTIONS OF CENTROSOME PROTEINS 11
2.1 CHOOSING TRANSGENIC CONSTRUCTS FOR THE SUBSEQUENT ANALYSIS .... 11
2.2 SPD-5 AND SPD-2 INTERACT IN CENTROSOME CONTAINING EXTRACT .... 14
2.3 IDENTIFICATION OF POSSIBLE INTERACTORS OF SPD-2 AND SPD-5 16
2.4 CHARACTERIZATION OF THE SHAPE AND MOLECULAR WEIGHT OF
SPD-5 AND SPD-2 20
2.4.1 SPD-2 EXISTS AS A MONOMER INSIDE THE CYTOPLASM 20
2.4.2 SPD-5 EXISTS MOSTLY AS A MONOMER BUT CAN ALSO FORM A
HETERO-TRIMER WITH RSA-L/RSA-2 IN THE CYTOPLASM 22
2.5 IN VIVO CHARACTERIZAION OF THE CYTOPLASMIC STATE OF SPD-2 AND SPD-5
26
2.5.1 FLUORESCENCE MICROSCOPY TECHNIQUES USED TO STUDY PROTEINS
IN VIVO
26
2.5.2 FUORESCENCE CORRELATION SPECTROSCOPY REVEALS DISTINCT DIFFU
SIVE BEHAVIOR OF GFP::SPD-2, SPD-5::GFP AND LAP::RSA-1 . 28
2.5.3 LAP::RSA-1 INTERACTS WITH RSA-2 AND SPD-5 IN THE CYTO
PLASM OF LIVING C. ELEGANS EMBRYOS 30
III
HTTP://D-NB.INFO/1070321206
2.5.4 THE DIFFUSION OF SPD-5::GFP DOES NOT CHANGE UPON DEPLE
TION OF RSA-2 31
2.5.5 THE DIFFUSION OF SPD-5 IS NOT DYNAMICALLY REGULATED
THROUGHOUT THE CELL CYCLE AND INDEPENDENT OF THE ANTERIOR-
POSTERIOR AXIS 31
2.5.6 SPD-2 DOES NOT INTERACT WITH SPD-5 IN THE CYTOPLASM OF LIV
ING C. ELEGANS EMBRYOS 36
3 DISCUSSION - CYTOPLASMIC INTERACTIONS OF CENTROSOME PROTEINS 38
3.1 ADVANTAGES AND LIMITATIONS OF THE USED TECHNIQUES 39
3.2 CHARACTERIZATION OF CYTOPLASMIC SPD-5 41
3.3 CHARACTERIZATION OF CYTOPLASMIC SPD-2 43
3.4 POSSIBLE CONSEQUENCES FOR PCM ASSEMBLY IN C. ELEGANS 44
4 RESULTS - A MECHANISM OF PCM ASSEMBLY IN C.
ELEGANS
46
4.1 SPD-5 IS HIGHLY PHOSPHORYLATED 46
4.2 PHOSPHORYLATION OF SPD-5 IS ESSENTIAL FOR PCM ASSEMBLY 48
4.3 PHOSPHORYLATION OF SPD-5 DOES NOT AFFECT ITS CYTOPLASMIC DIFFUSION
52
5 DISCUSSION - A MECHANISM OF PCM ASSEMBLY IN C.
ELEGANS
54
5.1 PHOSPHORYLATION OF SPD-5 DRIVES PCM ASSEMBLY IN C. ELEGANS ... 54
5.2 A UNIFIED MODEL FOR THE PCM CYCLE 57
5.2.1 BEHAVIOR OF SCAFFOLD PROTEINS PRIOR TO ASSEMBLY 59
5.2.2 PCM EXPANSION, CENTROSOME MATURATION AND FINAL CENTRO
SOME SIZE 59
5.2.3 PCM DISASSEMBLY DURING MITOTIC EXIT 60
5.3 GENERAL RELEVANCE OF THIS STUDY 60
6 MATERIALS AND METHODS 62
6.1 MOLECULAR CLONING AND MUTAGENESIS 62
6.2 WORM STRAINS 63
6.3 RNA INTERFERENCE 65
6.4 SCORING OF EMBRYO VIABILITY 66
6.5 IMMUNO BLOT ANALYSIS 66
6.6 PREPARATION OF LARGE MASSES OF EMBRYOS 67
6.7 PREPARATION OF EMBRYONIC EXTRACT 68
6.8 SIZE EXCLUSION CHROMATOGRAPHY AND RATE ZONAL ULTRACENTRIFUGATION
ANALYSIS 68
6.9 MOLECULAR WEIGHT AND SHAPE ANALYSIS 69
6.10 PREPARATION OF ANTIBODY COUPLED BEADS AND IMMUNO PRECIPITATION 69
6.11 LABEL FREE QUANTIFICATION MASS SPECTROMETRY ANALYSIS 70
6.12 MAPPING OF SPD-5 PHOSPHORYLATION SITES 71
6.13 PLK-1 PHOSPHORYLATION SITE PREDICTION 73
6.14 LIVE CELL IMAGING FOR CENTROSOME SIZE AND SPINDLE SIZE
DETERMINATION 73
6.15 FLUORESCENCE CORRELATION SPECTROSCOPY (FCS) AND DIFFUSION ANALYSIS
73
REFERENCES 76
A APPENDIX 87
A.L COMPLETE LIST OF PHOSPHORYLATION SITES IDENTIFIED ON SPD-5 IN VIVO
USING MASS SPECTROMETRY ANALYSIS 88
A.2 POTENTIAL PLK-1 PHOSPHORYLATION SITES PREDICTED FOR SPD-5 .... 89
A.3 AIR-1 RNAI DOES NOT CHANGE THE DIFFUSION OF SPD-5::GFP OR
GFP::SPD-2 90
A.4 EXAMPLARY FITS OF TYPICAL FCS MEASUREMENTS 91
A.5 EVALUATION OF THE GOODNESS OF THE FIT 92
A.6 OPTIMAL MEASUREMENT DEPTH FOR FCS MEASUREMENTS IN C. ELEGANS
EMBRYOS 93
ACKNOWLEDGEMENTS 95
DECLARATION 96
|
| any_adam_object | 1 |
| author | Wüseke, Oliver 1984- |
| author_GND | (DE-588)1069429228 |
| author_facet | Wüseke, Oliver 1984- |
| author_role | aut |
| author_sort | Wüseke, Oliver 1984- |
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| building | Verbundindex |
| bvnumber | BV042685771 |
| classification_rvk | WD 5100 |
| ctrlnum | (DE-599)DNB1070321206 |
| dewey-full | 571.84436541257 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.84436541257 |
| dewey-search | 571.84436541257 |
| dewey-sort | 3571.84436541257 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| indexdate | 2024-07-10T07:07:24Z |
| institution | BVB |
| language | English |
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| physical | VIII, 93, 2 S. Ill., graph. Darst. CD-ROM (12 cm) |
| publishDate | 2015 |
| publishDateSearch | 2015 |
| publishDateSort | 2015 |
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| spelling | Wüseke, Oliver 1984- Verfasser (DE-588)1069429228 aut Pericentriolar material assembly in Caenorhabditis elegans von Oliver Wüseke 2015 VIII, 93, 2 S. Ill., graph. Darst. CD-ROM (12 cm) txt rdacontent n rdamedia nc rdacarrier Dresden, Techn. Univ., Diss., 2015 Centrosom (DE-588)4295334-0 gnd rswk-swf Caenorhabditis elegans (DE-588)4147127-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Caenorhabditis elegans (DE-588)4147127-1 s Centrosom (DE-588)4295334-0 s DE-188 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028117541&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Wüseke, Oliver 1984- Pericentriolar material assembly in Caenorhabditis elegans Centrosom (DE-588)4295334-0 gnd Caenorhabditis elegans (DE-588)4147127-1 gnd |
| subject_GND | (DE-588)4295334-0 (DE-588)4147127-1 (DE-588)4113937-9 |
| title | Pericentriolar material assembly in Caenorhabditis elegans |
| title_auth | Pericentriolar material assembly in Caenorhabditis elegans |
| title_exact_search | Pericentriolar material assembly in Caenorhabditis elegans |
| title_full | Pericentriolar material assembly in Caenorhabditis elegans von Oliver Wüseke |
| title_fullStr | Pericentriolar material assembly in Caenorhabditis elegans von Oliver Wüseke |
| title_full_unstemmed | Pericentriolar material assembly in Caenorhabditis elegans von Oliver Wüseke |
| title_short | Pericentriolar material assembly in Caenorhabditis elegans |
| title_sort | pericentriolar material assembly in caenorhabditis elegans |
| topic | Centrosom (DE-588)4295334-0 gnd Caenorhabditis elegans (DE-588)4147127-1 gnd |
| topic_facet | Centrosom Caenorhabditis elegans Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028117541&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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