Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2001
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. Sprache. - Würzburg, Univ., Diss., 2001 |
| Beschreibung: | 114 Bl. Ill., graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Dinev, Dragomir |d 1970- |e Verfasser |0 (DE-588)123710294 |4 aut | |
| 245 | 1 | 0 | |a Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells |c Dragomir Dinev |
| 264 | 1 | |c 2001 | |
| 300 | |a 114 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Zsfassung in dt. Sprache. - Würzburg, Univ., Diss., 2001 | ||
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Datensatz im Suchindex
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TABLE OF CONTENTS
TABLE OF CONTENTS
I SUMMARY. 7
I ZUSAMMENFASSUNG. 8
II INTRODUCTION. 9
11-1. MAPK SIGNALLING PATHWAYS. 9
11-1.1. THE RAF/MEK/ERK MODULE. 10
11-1.2. THE STRESS-ACTIVATED JNK- AND P38 MAP KINASE CASCADES. 12
11-1.2.1. JNK-MAP KINASE CASCADE. 12
11-1.2.2. P38 MAP KINASE CASCADE. 13
11-1.2.3. PHYSIOLOGICAL ROLES OF JNK-AND P38 MAPK CASCADES. 13
11-1.3. MEK5/ERK5 SIGNALLING PATHWAY. 15
11-1.3.1 MEK5 - IDENTIFICATION, STRUCTURE AND FUNCTION. 15
11-1.3.2. ERK5- IDENTIFICATION, STRUCTURE, AND FUNCTION. 18
II-2. MEF2 FAMILY OF TRANSCRIPTION FACTORS. 23
11-2.1. STRUCTURE OF THE MEF2 FAMILY MEMBERS. 23
II-2.2. TISSUE AND CELL TYPE DISTRIBUTION OF MEF2 PROTEINS. 24
II-2.3. REGULATION OF MEF2 ACTIVITY. 25
11-2.3.1. TRANSCRIPTIONAL CONTROL. 25
II-2.3.2. TRANSLATIONAL CONTROL. 26
II-2.3.3. POST- TRANSLATIONAL CONTROL. 26
11-2.3.3.1. REGULATION BY PROTEIN INTERACTIONS. 26
II-2.3.3.2. REGULATION OF MEF2 PROTEINS VIA PHOSPHORYLATION. 27
II-2.3.4. CONTROL OF MEF2 TRANSCRIPTIONAL ACTIVITY BY INTERACTION WITH
MYOGENIC
BHLH PROTEINS. 27
II-2.3.5. A TWO-STEP MODEL FOR TRANSCRIPTIONAL ACTIVATION OF MUSCLE
SPECIFIC
1
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/964366584
TABLE OF CONTENTS
GENES. 29
11-3. SUPERFAMILY OF BASIC HELIX-LOOP-HELIX PROTEINS. 31
11-3.1. MYOGENIC REGULATORY FACTORS. 31
11-3.1.1 STRUCTURE OF MRFS. 31
II- 4. MYOGENIC DIFFERENTIATION. 34
11-4.1. MOUSE KNOCK-OUT STUDIES AND IN VITRO MYOGENIC DIFFERENTIATION.
34
II-4.2. REGULATION OF MYOGENESIS DURING DEVELOPMENT. 35
II- 4.3. EXTRACELLULAR HINTS THAT MEDIATE MYOGENIC DIFFERENTIATION. 35
IL-4.4. THE ROLE OF SIGNAL TRANSDUCTION IN MYOGENIC DIFFERENTIATION. 37
11-4.4.1. PROTEIN KINASE C (PKC). 37
II-4.4.2. THE MAPK SIGNALLING PATHWAY. 37
II-4.4.3. THE PHOSPHATIDYLINOSITOL 3-KINASE (PI3'K) PATHWAY. 38
AIM OF THE PROJECT. 40
III RESULTS. 41
III- 1. ERK5 BECOMES STRONGLY ACTIVATED UPON INDUCTION OF
DIFFERENTIATION
IN
C
2
C
12
MYOBLASTS. 41
III-2. TRANSACTIVATION OF DIFFERENTIATION SPECIFIC PROMOTERS IS
DEPENDENT ON
ERK5 ACTIVITY. 44
III- 2.1. MEK5/ERK5 UPREGULATE THE ACTIVITY OF THE P21 PROMOTER UNDER
DIFFERENTIATION CONDITIONS. 44
III-2.2. A 4XE-BOX CONTAINING PROMOTER IS STRONGLY TRANSACTIVATED BY
MEK5/ERK5 WHEN MUSCLE DIFFERENTIATION IS INDUCED. 45
III-2.3. MLC1A PROMOTER ACTIVITY IS INDUCED BY MEK5/ERK5 SIGNALLING
PATHWAY UNDER DIFFERENTIATION SPECIFIC CONDITIONS. 46
HI-3. ACTIVATION OF THE MEK5/ERK5 SIGNALLING PATHWAY UNDER
2
TABLE OF CONTENTS
DIFFERENTIATION CONDITIONS SUGGESTS A SYNERGISM BETWEEN MEF2C
AND MYOD ON TRANSCRIPTIONAL LEVEL. 48
111-3.1. MEK5/ERK5 DIFFERENTIATION MEDIATED ACTIVATION LEADS TO
UPREGULATION OF THE TRANSCRIPTIONAL ACTIVITY OF MEF2C. 48
III-3.2. ACTIVATED ERK IS ABLE TO PHOSPHORYLATE MYOD
IN VITRO. 49
ILL-4. BLOCK OF ERK5 EXPRESSION LEADS TO INHIBITION OF MYOGENIC
DIFFERENTIATION. 50
111-4.1. GENERATION OF STABLY EXPRESSING C
2C12 CELL LINES USING RETROVIRAL
INFECTION. 50
III-4.2. MYOGENIC DIFFERENTIATION IS BLOCKED WHEN THE EXPRESSION OF ERK5
IS
INHIBITED. 50
III-4.3. BLOCKAGE OF THE ENDOGENOUS ERK5 LEADS TO INHIBITION OF THE
EXPRESSION OF MYOGENIC PROTEINS. 51
III- 4.4. ERK5 OVEREXPRESSION IS SUFFICIENT FOR THE COMMITMENT OF C
2C12 CELLS
TO DIFFERENTIATE. 52
IV DISCUSSION. 55
IV-1. ERK5 BECOMES STRONGLY ACTIVATED UPON INDUCTION OF DIFFERENTIATION
IN C
2C12 MYOBLASTS. 55
IV-2. TRANSACTIVATION OF DIFFERENTIALLY-SPECIFIC PROMOTERS IS DEPENDENT
ON
ERK5 FUNCTIONAL ACTIVITY. 56
IV- 2.1. MEK5/ERK5 UPREGULATE THE ACTIVITY OF THE ENDOGENOUS P21
PROMOTER UNDER DIFFERENTIATION CONDITIONS. 57
IV-2.2. A EXPRESSION OF 4XE-BOX CONTAINING PROMOTER IS UPREGULATED BY
MEK5/ERK5 WHEN MUSCLE DIFFERENTIATION IS INDUCED. 58
IV-2.3. MLC1A IS INDUCED BY MEK5/ERK5 SIGNALLING PATHWAY UNDER
3
TABLE OF CONTENTS
DIFFERENTIATION SPECIFIC CONDITIONS. 59
IV-3. ACTIVATION OF THE MEK5/ERK5 SIGNALLING PATHWAY UNDER
DIFFERENTIATION CONDITIONS SUGGESTS A SYNERGISM BETWEEN MEF2C
AND MYOD ON TRANSCRIPTIONAL LEVEL. 60
IV-3.1. MEK5/ERK5 DIFFERENTIATION MEDIATED ACTIVATION LEADS TO
UPREGULATION OF THE TRANSCRIPTIONAL ACTIVITY OF MEF2C. 60
IV-3.2. ACTIVATED ERK IS ABLE TO PHOSPHORYLATE MYOD
IN VITRO. 61
IV- 4. BLOCK OF ERK5 EXPRESSION LEADS TO INHIBITION OF MYOGENIC
DIFFERENTIATION. 62
IV-4.1. GENERATION OF STABLY EXPRESSING C
2C12 CELL LINES. 62
IV-4.2. MYOGENIC DIFFERENTIATION IS BLOCKED WHEN THE EXPRESSION OF ERK5
IS
INHIBITED. 62
IV-4.3. BLOCKADE OF THE ENDOGENOUS ERK5 LEADS TO INHIBITION ON THE
MYOGENIC MARKERS. 63
IV- 4.4. ERK5 OVEREXPRESSION IS SUFFICIENT FOR THE COMMITMENT OF C
2C12 CELLS
TO DIFFERENTIATE. 64
V MATERIALS AND METHODS. 66
V- 1. MATERIALS. 66
V- 1.1. INSTRUMENTS. 66
V-1.2. REAGENTS AND GENERAL MATERIALS. 67
V-1.3. CELL CULTURE MATERIALS. 70
V-1.4. ENZYMES.
70
V-1.5. ANTIBODY CONJUGATES. 71
V-1.6. KITS. 71
V-1.7. PLASMIDS. 72
4
TABLE O F CONTENTS
V-1.8. PCR-PRIMERS. 73
V-1.9. BACTERIA STRAINS AND CELL LINES. 73
V-1.10. MEDIA. 73
V-1.11. SOLUTIONS. 74
V-2. METHODS. 80
V-2.1. WORKING WITH BACTERIA. 80
V.2.1.1. BACTERIAL CULTURE. 80
V-2.1.2. PREPARATION OF COMPETENT CELLS (CACI
2 METHOD). 80
V-1.2.3. TRANSFORMATION OF COMPETENT BACTERIA. 81
V-2.2. WORKING WITH DNA'S. 82
V-2.2.1. ELECTROPHORESIS OF DNA ON AGAROSE GEL. 82
V-2.2.2. ISOLATION OF PLASMID DNA FROM AGAROSE (QIAEX II AGAROSE
EXTRACTION
GEL PROTOCOL). 82
V-2.2.3. PURIFICATION OF PLASMID DNA (QIAQUICK PCR PURIFICATION KIT). 83
V-2.2.4. LIGATION OF DNA FRAGMENTS. 83
V-2.2.5. COHESIVE-END LIGATION. 84
V-2.2.6. MINI-PREPARATION OF PLASMID DNA. 84
V-2.2.7. MAXI-PREPARATION OF PLASMID DNA. 85
V-2.2.8. MEASUREMENT OF DNA CONCENTRATION. 85
V-2.2.9. DNA SEQUENCING (SANGER DIDEOXY METHOD). 86
V-2.3. WORKING WITH PROTEINS. 88
V-2.3.1. IMMUNOPRECIPITATION. 88
V-2.3.2.
IN VITRO KINASE ASSAYS. 89
V-2.3.3. MEASUREMENT OF PROTEIN CONCENTRATION (BIO-RAD PROTEIN ASSAY).
89
V-2.3.4. SODIUM DODECYL SULPHATE POLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-
5
TABLE OF CONTENTS
V-2.3.5.
V-2.4.
V-2.4.1.
V-2.4.2.
V-2.4.3.
V-2.4.4.
V-2.4.5.
V-2.5.
V-2.5.1.
V-2.5.2.
VI
VII
PAGE).
IMMUNOBLOTTING.
WORKING WITH CELL CULTURES.YY
MAINTENANCE OF CELL CULTURE.
CALCIUM PHOSPHATE TRANSIENT TRANSFECTION ASSAY.
LIPOFECTAMINE 2000
YY
TRANSIENT TRANSFECTION ASSAY.
LUCIFERASE REPORTER GENE ASSAY.
P-GALACTOSIDASE ASSAY.
RETROVIRAL INFECTIONS.
TRANSIENT TRANSFECTION FOR PRODUCTION OF INFECTIOUS RETROVIRUSES.
PRODUCTION OF STABLY TRANSDUCED C
2C12 CELL LINES USING RETROVIRAL
INFECTIONS.
REFERENCES.
APPENDIX.
89
91
91
91
92
93
93
94
95
95
95
96
111
6 |
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| spelling | Dinev, Dragomir 1970- Verfasser (DE-588)123710294 aut Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells Dragomir Dinev 2001 114 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. Sprache. - Würzburg, Univ., Diss., 2001 Zelldifferenzierung (DE-588)4067536-1 gnd rswk-swf Muskelzelle (DE-588)4127323-0 gnd rswk-swf Proteinkinasen (DE-588)4175996-5 gnd rswk-swf Signaltransduktion (DE-588)4318717-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Muskelzelle (DE-588)4127323-0 s Zelldifferenzierung (DE-588)4067536-1 s Signaltransduktion (DE-588)4318717-1 s Proteinkinasen (DE-588)4175996-5 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009673286&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Dinev, Dragomir 1970- Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells Zelldifferenzierung (DE-588)4067536-1 gnd Muskelzelle (DE-588)4127323-0 gnd Proteinkinasen (DE-588)4175996-5 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| subject_GND | (DE-588)4067536-1 (DE-588)4127323-0 (DE-588)4175996-5 (DE-588)4318717-1 (DE-588)4113937-9 |
| title | Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells |
| title_auth | Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells |
| title_exact_search | Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells |
| title_full | Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells Dragomir Dinev |
| title_fullStr | Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells Dragomir Dinev |
| title_full_unstemmed | Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells Dragomir Dinev |
| title_short | Analysis of the role of extracellular signal regulated kinase 5 (ERK5) in the differentiation of muscle cells |
| title_sort | analysis of the role of extracellular signal regulated kinase 5 erk5 in the differentiation of muscle cells |
| topic | Zelldifferenzierung (DE-588)4067536-1 gnd Muskelzelle (DE-588)4127323-0 gnd Proteinkinasen (DE-588)4175996-5 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| topic_facet | Zelldifferenzierung Muskelzelle Proteinkinasen Signaltransduktion Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009673286&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT dinevdragomir analysisoftheroleofextracellularsignalregulatedkinase5erk5inthedifferentiationofmusclecells |