Regulation of plant potassium channels:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2015
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XIV, 115 S. Ill., graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Skłodowski, Kamil |e Verfasser |0 (DE-588)1076545254 |4 aut | |
| 245 | 1 | 0 | |a Regulation of plant potassium channels |c von Kamil Skłodowski |
| 264 | 1 | |c 2015 | |
| 300 | |a XIV, 115 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
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| 502 | |a Potsdam, Univ., Diss., 2015 | ||
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Datensatz im Suchindex
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|---|---|
| adam_text | CONTENTS
CONTENTS II
ACKNOWLEDGEMENTS VII
DECLARATION VIII
ABSTRACT IX
DIE ZUSAMMENFASSUNG XI
LIST OF ABBREVIATIONS XII
1 INTRODUCTION 1
1.1 PLANT VOLTAGE GATED POTASSIUM CHANNELS 1
1.1.1 K
+
AND ITS ROLE IN PLANTS 1
1.1.2 TRANSPORT OF POTASSIUM IN PLANTS 2
1.1.3 ARABIDOPSIS AKT2 CHANNEL 4
1.2 PLANT KINASES 6
1.2.1 RLK IN PLANTS 6
1.2.2 ROLE OF LRU RLK IN PLANT LIFE 7
1.3 SUCROSE AND ITS ROLE IN PLANTS 8
1.3.1 SOURCE TO SINK - LONG DISTANCE SUCROSE TRANSPORT 8
1.3.2 ROLE OF SUCROSE IN SIGNALLING 10
1.3.3 LINKS BETWEEN CHANNELS, KINASES AND SUCROSE 11
1.3.4 AIM OF THE RESEARCH 13
2 MATERIALS AND METHODS 15
2.1 MATERIALS 15
2.1.1 KITS 15
2.1.2 VECTORS 16
2.1.3 BIOLOGICAL MATERIAL 16
2.1.4 PRIMERS 17
2.1.5 ENZYMES 18
2.1.6 PURCHASED MEDIA, BUFFERS AND OTHERS 18
2.1.7 NON-PURCHASED MEDIA, BUFFERS AND OTHERS 19
2.2 METHODS 20
2.2.1 CONSTRUCT PREPARATION 20
2.2.2 DNA EXTRACTION AND PURIFICATION 20
III
HTTP://D-NB.INFO/107784168X
2.2.3 PROTEIN EXTRACTION 21
2.2.4 SDS-PAGE AND WESTERN BLOTTING 21
2.2.5 GROWTH OF PLANTS 21
2.2.6 GROWTH OF MICROORGANISMS 22
2.2.7 Y2H SCREENING 22
2.2.8 SPLIT UBIQUITIN SYSTEM 23
2.2.9 PROTOPLAST ISOLATION 24
2.2.10 GOLD PARTICLE COATING AND BOMBARDMENT 24
2.2.11 TWO-ELECTRODE VOLTAGE-CLAMP (TEVC) 25
2.2.12 FLUORESCENT MICROSCOPY 26
2.2.13 ROOT HAIR ANALYSIS 28
3 RESULTS 31
3.1 THE K
+
BATTERY-REGULATING ARABIDOPSIS K
+
CHANNEL AKT2 IS UNDER THE
CONTROL OF MULTIPLE POST-TRANSLATIONAL STEPS 31
3.1.1 SYNOPSIS 31
3.1.2 AUTHOR S CONTRIBUTION 32
3.1.3 ORIGINAL PUBLICATION 32
3.2 SCREENING FOR PUTATIVE INTERACTION PARTNERS USING YEAST-2-HYBRID
SYSTEM . 38
3.2.1 SYSTEM OPTIMIZATION 38
3.2.2 SCREENING AND DATA FILTERING 40
3.2.3 1-ON-L VERIFICATION OF THE SELECTED CANDIDATES USING
YEAST-2-HYBRID
SYSTEM 42
3.3 PHENOTYING MRHL KNOCK-OUT PLANTS FOR SIMILARITIES TO AKT2 KNOCK-OUT
PLANTS 44
3.3.1 ANALYSIS OF STEM LENGTH 45
3.3.2 TESTING THE NEW SEMI-AUTOMATED METHOD FOR THE ANALYSIS OF ROOT
HAIR FORMATION 47
3.3.3 ANALYSIS OF ROOT HAIR FORMATION IN AKT2-L AND MRHL-1 MUTANT PLANTS
49
3.4 SUBCELLULAR LOCALISATION AND FRET ANALYSIS OF AKT2 AND ITS PUTATIVE
INTERACTING PARTNERS 52
3.4.1 LOCALISATION OF AKT2 SUBUNITS 53
3.4.2 CO-LOCALISATION OF AKT2 SUBUNITS WITH KAT2, MRH1 AND MRH2 . 55
3.4.3 ANALYSIS OF POSSIBLE AKT2 AND MRH1 INTERACTION IN ARABIDOPSIS
PROTOPLASTS BY FRET 59
3.5 ANALYSIS OF AKT2 RESIDUES RESPONSIBLE FOR POSSIBLE KINASE DOCKING 61
3.5.1 MOLECULAR DYNAMICS ANALYSIS OF POSSIBLE DOCKING SITES 62
3.5.2 FUNCTIONAL ANALYSIS OF AKT2 VARIANTS IN XENOPUS LAEVIS OOCYTES
BY TEVC 63
3.6 TESTING AKT2 AND ITS VARIANTS IN THE SPLIT UBIQUITIN SYSTEM 64
3.6.1 VERIFICATION OF SYSTEM FUNCTIONALITY 65
3.6.2 TESTING AKT2 VARIANTS IN COMBINATION WITH MRH1 AND MRH2 IN
SUS 66
3.7 THE EFFECT OF MRH1 ON AKT2 FUNCTIONALITY COMBINED WITH BIFC ANALYSIS
69
3.8 DETAILED ANALYSIS OF THE CYTOSOLIC PARTS OF AKT2 EXPLAINS ITS POOR
MEM
BRANE LOCALISATION IN PLANT CELLS 72
3.8.1 TESTING THE EFFECT OF C-TERMINALLY FUSED FLUOROCHROME ON AKT2
INTERACTION WITH KAT2 IN SUS 72
IV
3.8.2 TESTING IN PLANTA LOCALISATION AND FUNCTIONALITY OF THE
ALTERNATIVE
YFP FUSIONS WITH AKT2 75
4 DISCUSSION 79
4.1 REGULATION OF VOLTAGE GATED PLANT POTASSIUM CHANNELS 79
4.1.1 REGULATION OF AKT2 CHANNEL VIA PHOSPHORYLATION 80
4.1.1.1 IMPORTANCE OF AKT2 GATING MODES IN ABOVEGROUND AND
UNDERGROUND PARTS OF THE PLANT 81
4.1.1.2 ADDITIONAL ANALYSIS OF ROOT HAIR FORMATION USING A NEW
TOOL REVEALED DIFFERENCES IN THE ROOT HAIR ARCHITECTURE ... 84
4.1.1.3 MOLECULAR ANALYSIS OF THE INTERACTION BETWEEN AKT2 AND
THE MRH1 SHOWED HIGH COMPLEXITY OF THE AKT2 REGULATION 86
4.1.1.4 DETAILED ANALYSIS OF THE PUTATIVE AKT2 PHOSPHORYLATION
SITES INDICATES A TRANSIENT NATURE OF AKT2-MRH1 INTER
ACTION 88
4.1.1.5 EFFECTS OF MRH1 ON AKT2 FUNCTION ARE VISIBLE ONLY WHEN
THE BIFC SYSTEM IS EMPLOYED 90
4.2 THE C-TERMINUS OF AKT2 IS ESSENTIAL FOR CHANNEL TETRAMERIZATION 91
4.2.1 LARGE C-TERMINAL FUSIONS DETAIN THE CHANNEL IN THE INTERMEDIATE
LOCALISATION 92
4.2.2 ALTERNATIVE FLUOROPHORE FUSION SITES YIELD FUNCTIONAL CHANNELS AND
CORRECT LOCALIZATION IN PLANT CELLS 93
4.3 SUMMARY 94
5 CONCLUSIONS 97
APPENDIX A 99
APPENDIX B 100
APPENDIX C 101
BIBLIOGRAPHY 105
V
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| bvnumber | BV042890916 |
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| indexdate | 2024-07-10T07:12:11Z |
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| language | English |
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| physical | XIV, 115 S. Ill., graph. Darst. |
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| spelling | Skłodowski, Kamil Verfasser (DE-588)1076545254 aut Regulation of plant potassium channels von Kamil Skłodowski 2015 XIV, 115 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Potsdam, Univ., Diss., 2015 (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=028319417&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Skłodowski, Kamil Regulation of plant potassium channels |
| subject_GND | (DE-588)4113937-9 |
| title | Regulation of plant potassium channels |
| title_auth | Regulation of plant potassium channels |
| title_exact_search | Regulation of plant potassium channels |
| title_full | Regulation of plant potassium channels von Kamil Skłodowski |
| title_fullStr | Regulation of plant potassium channels von Kamil Skłodowski |
| title_full_unstemmed | Regulation of plant potassium channels von Kamil Skłodowski |
| title_short | Regulation of plant potassium channels |
| title_sort | regulation of plant potassium channels |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028319417&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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