Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Erlangen ; Nürnberg
[2018]
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | 99 Seiten Illustrationen, Diagramme 21 cm |
Internformat
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| 245 | 1 | 0 | |a Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway |c vorgelegt von Kowcee Jalal |
| 246 | 1 | 3 | |a Functional characterisation of PGAM5 in the WNT/beta-catenin signalling pathway |
| 264 | 1 | |a Erlangen ; Nürnberg |c [2018] | |
| 300 | |a 99 Seiten |b Illustrationen, Diagramme |c 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |b Dissertation |c Friedrich-Alexander-Universität Erlangen-Nürnberg |d 2018 | ||
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Datensatz im Suchindex
| _version_ | 1804178387444432896 |
|---|---|
| adam_text | CONTENTS
ZUSAMMENFASSUNG
.................
7
SUMMARY.....................................................................................................................
8
INTRODUCTION............................................................................................................
9
THE WNT SIGNALLING PATHWAY
...................................................................................
9
THE WNT/P-CATENIN SIGNAL
TRANSDUCTION........................................................................10
COMPONENTS OF THE WNT/SS-CATENIN SIGNALLING PATHWAY
............................
12
WNT
LIGANDS.....................................................................................................................12
PORCUPINE........................................................................................................................12
FRIZZLED
RECEPTORS..........................................................................................................13
LOW DENSITY LIPOPROTEIN (LDL) RECEPTOR RELATED PROTEINS
..........................................
13
DISHEVELLED.....................................................................................................................13
GLYCOGEN SYNTHASE KINASE 3 BETA
................................................................................
13
CASEIN KINASE
1..............................................................................................................14
ADENOMATOUS POLYPOSIS
COLI........................................................................................14
AXINL.....................................................................................
14
SS-CATENIN..........................................................................................................................15
T-CELL FACTOR/LYMPHOID ENHANCER BINDING FACTOR
....................................................
15
PHOSPHOGLYCERATE MUTASE
5..................................................................................16
PGAM5 AND THE WNT/SS-CATENIN SIGNALLING PATHWAY
......................................
22
RESULTS.....................................................................................................................
23
ANALYSIS OF THE PGAM5 PROTEIN IN THE CELL LINE 293T
...............
23
LOCALISATION OF
PGAM5.................................................................................................
25
INTERACTION PARTNERS OF
PGAM5....................................................................................
27
PGAM5 INTERACTS WITH
AXINL.........................................................................................
27
CO-LOCALISATION OF PGAM5 AND A XIN
L..........................................................................
29
FUNCTION OF PGAM5L IN THE WNT/SS-CATENIN SIGNALLING PATHWAY
...............
31
EFFECT OF PGAM5L OVEREXPRESSION ON THE WNT/SS-CATENIN SIGNALLING PATHWAY
........
31
PGAM5L EXERTS ITS EFFECTS VIA SS-CATENIN ON THE WNT/SS-CATENIN SIGNALLING
PATHWAY33
PGAM5L OVEREXPRESSION INCREASES SS-CATENIN LEVELS
................................................
35
EFFECT OF TRUNCATED (CYTOPLASMIC) PGAM5L OVEREXPRESSION ON THE
WNT/SS-CATENIN
SIGNALLING
PATHWAY........................................................................................................
37
EFFECT OF THE N-TERMINALLY TAGGED PHOSPHATASE DEAD PGAM5L MUTANT
ON THE WNT/SS-CATENIN SIGNALLING PATHWAY
.......................................................
39
THE PHOSPHATASE MOTIF OF PGAM5L IS NECESSARY FOR THE ACTIVATION OF THE
WNT/SS-
CATENIN SIGNALLING
PATHWAY...........................................................................................
39
N-TERMINALLY TAGGED PHOSPHATASE DEAD PGAM5L MUTANT DECREASES AXINL
EXPRESSION
LEVELS..........................................................................................................
39
PROCESSING AND FUNCTION OF ENDOGENOUS PGAM5L
.......................................
42
ENDOGENOUS CLEAVAGE OF PGAM5 AND RELEASE INTO CYTOPLASM
.................................
42
HYPOXIA CLEAVES ENDOGENOUS PGAM5 AND THIS LEADS TO ITS INTERACTION WITH
MEMBERS OF THE WNT/SS-CATENIN SIGNALLING PATHWAY
....................................................
43
EFFECTS OF LOSS OF FUNCTION OF PGAM5
.............................................................
46
SIRNA KNOCKDOWN OF
PGAM5.......................................................................................
46
GENERATION OF 293T HEK PGAM5 KNOCKOUT CELL LINES
................................................
48
293T PGAM5 KNOCKOUT CELL LINES AND
HYPOXIA............................................................48
SIRNA KNOCKDOWN OF SS-CATENIN INCREASES ENDOGENOUS PGAM5 EXPRESSION
LEVELS
..........................................................................................................................................
50
FUNCTIONAL ANALYSIS OF
PGAM5S............................................................................52
EFFECT OF PGAM5S OVEREXPRESSION ON THE WNT/SS-CATENIN SIGNALLING PATHWAY
........
52
EXPRESSION OF PGAM5S PROTEIN IN 293T
CELLS..............................................................53
ENDOGENOUS PGAM5S EXPRESSION IN 293T
CELLS.........................................................53
ENDOGENOUS PGAM5S EXPRESSION IN DIFFERENT CELL LINES
...........................................
54
DISCUSSION................................................................................................................56
INTERACTION ANALYSIS OF
PGAM5.............................................................................56
LOCALISATION ANALYSIS OF
PGAM5...........................................................................57
FUNCTIONAL ANALYSIS OF PGAM5 IN THE WNT/SS-CATENIN SIGNALLING
PATHWAY............................................................................................................................58
POSITIVE REGULATION OF THE WNT/SS-CATENIN SIGNALLING PATHWAY BY PGAM5
.................
58
NEGATIVE REGULATION OF THE WNT/SS-CATENIN SIGNALLING PATHWAY BY PGAM5
...............
59
PGAM5 AND THE PHOSPHATASE
MOTIF..............................................................................59
FUNCTIONAL ANALYSIS OF ENDOGENOUS PGAM5
...................................................
61
PGAM5-HYPOXIA-WNT/SS-CATENIN SIGNALLING PATHWAY AXIS
..........................................
61
THE PGAM5-SS-CATENIN SIGNALLING
PATHWAY.........................................................63
THE ALTERNATIVE SPLICE VARIANTS: PGAM5L AND PGAM5S
...............................
66
OUTLOOK............................................................................................................................68
MATERIALS..................................................................................................................70
USED CELL
LINES.............................................................................................................
70
CHEMICALS........................................................................................................................
70
SOLUTIONS AND
BUFFERS....................................................................................................
70
MOLECULAR BIOLOGY
REAGENTS...............................................................................72
OTHER
REAGENTS...........................................................................................................
72
MEDIUM FOR BACTERIAL
CELLS...................................................................................72
MEDIUM UND REAGENTS FOR EUKARYOTIC CELLS
.................................................
73
TRANSFECTION
REAGENTS............................................................................................
73
ANTIBODIES.......................................................................................................................
73
PRIMARY
ANTIBODIES........................................................................................................
73
SECONDARY
ANTIBODIES...................................................................................................
74
SYNTHETIC
OLIGONUCLEOTIDES..................................................................................74
SIRNAS.............................................................................................................................
74
PLASMIDS....................................................................................................................
74
CRISPR/CAS9 PGAM5
PLASMIDS..................................................................................
75
OTHER EQUIPMENT AND
SOFTWARE............................................................................76
METHODS.....................................................................................................................77
CELL CULTURE
METHODS..............................................................................................
77
PASSAGING OF EUKARYOTIC
CELLS.......................................................................................77
FREEZING AND THAWING OF EUKARYOTIC
CELLS....................................................................78
TRANSIENT TRANSFECTION OF EUKARYOTIC
CELLS...................................................................78
TRANSIENT TRANSFECTION WITH PEI (POLYETHYLENEIMINE) REAGENT
...................................
78
SILENCING OF GENE EXPRESSION BY RNA INTERFERENCE (RNAI)
.......................
79
PGAM5 AND SS-CATENIN SIRNA
KNOCKDOWN....................................................................79
PROTEIN
BIOCHEMISTRY................................................................................................80
PREPARATION OF CELL
LYSATES.............................................................................................80
DETERMINATION OF THE TOTAL PROTEIN QUANTITY IN CELL LYSATES /
RECOMBINANT PROTEINS 80
IMMUNOPRECIPITATION / CO-IMMUNOPRECIPITATION
.........................................................
80
SODIUM DODECYLSULFATE (SDS) POLYACRYLAMIDE GEL ELECTROPHORESIS
(PAGE) ACCORDING TO
LAEMMLI..................................................................................80
WESTERN
BLOT................................................................................................................81
IMMUNOFLUORESCENCE MICROSCOPY
......................................................................
82
IMMUNOFLUORESCENCE
STAINING......................................................................................82
REPORTER
ASSAYS.........................................................................................................83
REPORTER GENE MEASUREMENTS IN MAMMALIAN CELLS (TOP/FOPGLOW ASSAY)
...............
83
REPORTER GENE MEASUREMENTS IN MAMMALIAN CELLS (PBAR/RENILLA ASSAY)
..............
84
MOLECULAR BIOLOGY
METHODS.................................................................................84
CULTURE OF BACTERIAL
CELLS...............................................................................................85
HEAT SHOCK TRANSFORMATION OF COMPETENT E. COIL
........................................................
85
ISOLATION OF PLASMID DNA ON ANALYTICAL / PREPARATIVE SCALE
......................................
85
PHOTOMETRIC QUANTIFICATION OF NUCLEIC ACID CONCENTRATION OF THE
PURIFIED, ISOLATED
PLASMID
DNA...................................................................................................................86
HYPOXIA
ASSAY...............................................................................................................86
HYPOXIA
EXPOSURE..........................................................................................................87
REFERENCES.............................................................................................................
88
APPENDIX...................................................................................................................
98
ABBREVIATIONS........................................ 98
DIMENSIONS.........................................................................................................................98
UNITS....................................................................................................................................99
PUBLICATIONS...........................................................................................................
99
|
| any_adam_object | 1 |
| author | Jalal, Kowcee |
| author_facet | Jalal, Kowcee |
| author_role | aut |
| author_sort | Jalal, Kowcee |
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| building | Verbundindex |
| bvnumber | BV044862611 |
| ctrlnum | (OCoLC)1135390024 (DE-599)DNB1153795388 |
| discipline | Biologie |
| format | Thesis Book |
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| spelling | Jalal, Kowcee Verfasser aut Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway vorgelegt von Kowcee Jalal Functional characterisation of PGAM5 in the WNT/beta-catenin signalling pathway Erlangen ; Nürnberg [2018] 99 Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich-Alexander-Universität Erlangen-Nürnberg 2018 Signaltransduktion (DE-588)4318717-1 gnd rswk-swf Wnt-Proteine (DE-588)7515809-7 gnd rswk-swf Catenine (DE-588)4380415-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Signaltransduktion (DE-588)4318717-1 s Wnt-Proteine (DE-588)7515809-7 s Catenine (DE-588)4380415-9 s DE-604 B:DE-101 application/pdf http://d-nb.info/1153795388/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030257250&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Jalal, Kowcee Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway Signaltransduktion (DE-588)4318717-1 gnd Wnt-Proteine (DE-588)7515809-7 gnd Catenine (DE-588)4380415-9 gnd |
| subject_GND | (DE-588)4318717-1 (DE-588)7515809-7 (DE-588)4380415-9 (DE-588)4113937-9 |
| title | Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway |
| title_alt | Functional characterisation of PGAM5 in the WNT/beta-catenin signalling pathway |
| title_auth | Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway |
| title_exact_search | Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway |
| title_full | Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway vorgelegt von Kowcee Jalal |
| title_fullStr | Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway vorgelegt von Kowcee Jalal |
| title_full_unstemmed | Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway vorgelegt von Kowcee Jalal |
| title_short | Functional characterisation of PGAM5 in the WNT/β-catenin signalling pathway |
| title_sort | functional characterisation of pgam5 in the wnt β catenin signalling pathway |
| topic | Signaltransduktion (DE-588)4318717-1 gnd Wnt-Proteine (DE-588)7515809-7 gnd Catenine (DE-588)4380415-9 gnd |
| topic_facet | Signaltransduktion Wnt-Proteine Catenine Hochschulschrift |
| url | http://d-nb.info/1153795388/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030257250&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT jalalkowcee functionalcharacterisationofpgam5inthewntbcateninsignallingpathway AT jalalkowcee functionalcharacterisationofpgam5inthewntbetacateninsignallingpathway |
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