Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2014
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XIV, 113 S. Ill., graph. Darst. |
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Datensatz im Suchindex
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| adam_text | CONTENT
ABSTRACT III
ZUSAMMENFASSUNG V
FIGURE INDEX XI
ABBREVIATIONS XIII
1. INTRODUCTION 1
1.1. WNT SIGNALING 1
1.1.1. WNT/P-CATENIN SIGNALING: KEY COMPONENTS AND SIGNAL TRANSDUCTION 2
1.1.2. CELL CYCLE AND WNT SIGNALING 4
1.1.2.1. CELL CYCLE, CDK, AND CYCLIN -. 4
1.1.2.2. MITOGENIC AND MITOTIC WNT SIGNALING 6
1.2. WNT CO-RECEPTOR LRP6 8
1.2.1. LRP6 EXTRACELLULAR DOMAIN 8
1.2.2. LRP6 MATURATION 9
1.2.3. LRP6 INTRACELLULAR DOMAIN: PPPSP AND CK1 SITES 10
1.2.4. MODELS OF WNT RECEPTOR ACTIVATION 13
1.2.5. ENDOCYTOSIS IN WNT/LRP6 SIGNALING AND LRP6 TURNOVER 14
1.3. TYROSINE KINASES IN WNT SIGNALING 16
1.3.1. FER 17
1.3.2. SRC 18
1.3.3. FER AND SRC IN WNT/P-CATENIN SIGNALING 19
1.4. AIM OF STUDY 20
2. MATERIALS AND METHODS 21
2.1. MATERIALS 21
2.1.1. GROWTH MEDIA 21
2.1.2. BUFFER...: 21
2.1.3. ANTIBODIES 24
2.2. METHODS 24
2.2.1. CELL CULTURE 24
2.2.2. CELL TRANSFECTION AND HARVEST 25
2.2.3. TOPFLASH WNT LUCIFERASE REPORTER ASSAY 25
2.2.4. SDS-PAGE/WB 26
2.2.5. MEDAKA CDNA LIBRARY PREPARATION 28
VII
HTTP://D-NB.INFO/1048499057
2.2.6. MEDAKA CDNA SCREEN USING KINASE CDNA SUBSET 29
2.2.7. CELL CYCLE ARREST USING DOUBLE THYMIDINE BLOCK 29
2.2.8. IMMUNOFLUORESCENCE MICROSCOPY 30
2.2.9. IN VITRO SITE-DIRECTED MUTAGENESIS 30
2.2.10. IP/CO-IP 32
2.2.11
.IN
VITRO
KINASE ASSAY . 33
2.2.12. OPTIPREP DENSITY GRADIENT CENTRIFUGATION FOR PREPARATION OF DRM
FRACTIONS 33
2.2.13. OPTIPREP DENSITY GRADIENT CENTRIFUGATION FOR PREPARATION OF LRP6
AGGREGATES 34
3. RESULTS 35
3.1. PART I: CDK14/CYCLIN Y REGULATES LRP6 PHOSPHORYLATION 35
3.1.1. CDK14/CYCLINY IS A NOVEL PPPSP KINASE FOR LRP6 35
3.1.2. CYCLIN Y IS A MEMBRANE CYCLIN 36
3.1.3. CDK14/CYCLIN Y ASSOCIATES WITH LRP6 AND PHOSPHORYLATES LRP6 38
3.1.4. CELL CYCLE REGULATION OF LRP6 PHOSPHORYLATION 39
3.2. PART II: MEDAKA CDNA SCREEN FOR NEW MODIFIERS OF LRP6 AND CYCLIN Y
....42
3.2.1. MEDAKA CDNA LIBRARY PREPARATION 42
3.2.2. SCREEN FOR NEW MODIFIERS OF LRP6 AND CYCLIN Y USING MEDAKA KINASE
CDNA LIBRARY SUBSET 43
3.2.3. PKA PHOSPHORYLATES CYCLIN Y AND ACTIVATES WNT SIGNALING 47
3.3. PART III: TYROSINE PHOSPHORYLATION AND REGULATION OF LRP6 50
3.3.1. FER PHOSPHORYLATES LRP6 IN A KINASE DEPENDENT MANNER 50
3.3.2. FER INTERACTS WITH LRP6 AND DIRECTLY PHOSPHORYLATES LRP6 52
3.3.3. FER INHIBITS WNT/LRP6 SIGNALING 53
3.3.4. WNT3A STIMULATES LRP6 TYROSINE PHOSPHORYLATION BY FER 55
3.3.5. LRP6 PHOSPHORYLATION SITES FOR FER 57
3.3.6. INTERACTION BETWEEN LRP6 TYROSINE PHOSPHORYLATION BY FER AND
SERINE/THREONINE PHOSPHORYLATION AT PPPSP AND CK1 SITES 62
3.3.7. MECHANISM FOR FER/SRC INHIBITION ON LRP6 ACTIVATED WNT SIGNALING
66
3.3.7.1. FER ACCELERATES LRP6 TURNOVER 66
3.3.7.2. FER INCREASES IMMATURE FORM OF LRP6 67
3.3.7.3. REGULATION OF TYROSINE PHOSPHORYLATION ON LRP6 DISTRIBUTION
BETWEEN
LIPID AND NON-LIPID RAFTS 71
VIII
3.3.7.4. REGULATION OF TYROSINE PHOSPHORYLATION ON LRP6 SIGNALOSOME
FORMATION 74
4. DISCUSSION 81
4.1. PART I: CELL CYCLE REGULATION OF WNT RECEPTOR ACTIVATION 81
4.2. PART II: MEDAKA CDNA SCREEN FOR NEW MODIFIERS OF CYCLIN Y AND LRP6
....83
4.3. PART III: TYROSINE PHOSPHORYLATION AND REGULATION OF LRP6 85
4.3.1. CHARACTERIZATION OF FER MEDIATED LRP6 TYROSINE PHOSPHORYLATION 85
4.3.2. FER INHIBITS WNT/LRP6 SIGNALING 88
4.3.3. REGULATION OF FER MEDIATED LRP6 TYROSINE PHOSPHORYLATION 89
4.4.5. MECHANISM FOR FER/SRC MEDIATED INHIBITION FUNCTION ON WNT/LRP6
SIGNALING 91
4.4.5.1. FER FUNCTION ON LRP6 MATURATION AND TURNOVER 91
4.4.3.2. FER/SRC MEDIATED REGULATION ON LRP6 MEMBRANE DISTRIBUTION AND
SIGNALOSOME FORMATION 93
REFERENCES.- 95
IX
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| any_adam_object | 1 |
| author | Su, Yi |
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| bvnumber | BV041879735 |
| ctrlnum | (OCoLC)880388692 (DE-599)BSZ401291227 |
| dewey-full | 571.74 |
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| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.74 |
| dewey-search | 571.74 |
| dewey-sort | 3571.74 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
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| spelling | Su, Yi Verfasser (DE-588)1047571773 aut Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer von Yi Su 2014 XIV, 113 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Karlsruhe, KIT, Diss., 2014 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027323883&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Su, Yi Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer |
| subject_GND | (DE-588)4113937-9 |
| title | Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer |
| title_auth | Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer |
| title_exact_search | Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer |
| title_full | Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer von Yi Su |
| title_fullStr | Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer von Yi Su |
| title_full_unstemmed | Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer von Yi Su |
| title_short | Phosphorylation and regulation of the Wnt co-receptor LRP6 by Cyclin dependent kinase 14/Cyclin Y and Tyrosine kinase Fer |
| title_sort | phosphorylation and regulation of the wnt co receptor lrp6 by cyclin dependent kinase 14 cyclin y and tyrosine kinase fer |
| topic_facet | Hochschulschrift |
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