Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana:
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| Format: | Abschlussarbeit Mikrofilm Buch |
| Sprache: | English |
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2017
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| Beschreibung: | 2 Mikrofiches (XIII, 107 Seiten) Illustrationen, Diagramme |
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| 245 | 1 | 0 | |a Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana |c vorgelegt von Hemal Bhasin |
| 264 | 1 | |a Köln |c 2017 | |
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Datensatz im Suchindex
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| adam_text | TABLE OF CONTENTS
ACKNOWLEDGEMENT..........................................................................................................
I
ZUSAMMENFASSUNG.........................................................................................................ILL
ABSTRACT..............................................................................................................................V
PUBLICATION......................................................................................................................VII
ABBREVIATIONS..................................................................................................................IX
LIST OF
FIGURES..................................................................................................................XI
TABLES................................................................................................................................XIII
GENERAL
INTRODUCTION..................................................................................................1
ANGUSTIFOLIA: A CELL MORPHOGENESIS GENE WITH PLEIOTROPIC
EFFECTS.................................1
AN AND CTBP S : RELATED STRUCTURES BUT DISTINCT
FUNCTIONS?....................................................2
LOCALIZATION OF AN: A STORY OF GROWING
CONFUSION...............................................................4
AN EMS-SCREEN BASED NADH BINDING MUTANT OF
AN IS A NULL MUTANT
................................
4
AIMS OF MY
RESEARCH.......................................................................................................6
CHAPTER 1. AN IS A NOVEL COMPONENT OF MRNP
GRANULES..........................................................7
INTRODUCTION..................................................................................................................7
RNA BINDING PROTEINS: KEY REGULATORY MOLECULES INVOLVED IN
POST-TRANSCRIPTIONAL
CONTROL..................................................................................................................................7
MRNP GRANULES: MODULATORS OF GENE EXPRESSION IN RESPONSE TO STRESS
.............................
8
RESULTS.............................................................................................................................10
MALDI-MS ANALYSIS IDENTIFIED RNA BINDING PROTEINS THAT EXIST IN A
COMPLEX WITH
ANGUSTIFOLIA IN
VIVO..................................................................................................10
AN CO-LOCALIZES WITH KNOWN RNA BINDING PROTEINS TO MRNP GRANULES/SGS
....
...........
22
AN LOCALIZATION DYNAMICALLY CHANGES UPON THE APPLICATION OF HEAT AND
SALT STRESS IN AN-
YFP EXPRESSING TRANSGENIC
LINE........................................................................................26
DISCUSSION.......................................................................................................................30
AN IS A NOVEL COMPONENT OF SGS IN
ARABIDOPSIS.............................................................30
AN LOCALIZATION CHANGES DYNAMICALLY IN A STRESS DEPENDENT MANNER
..............................
31
AN IS INVOLVED IN POST-TRANSCRIPTIONAL/TRANSLATIONAL
REGULATION......................................32
AN MIGHT BE INVOLVED IN VESICLE MEDIATED TRANSPORT
.....................................................
32
CHAPTER 2. ROLE OF AN AS A COMPONENT OF 80S
.....................................................................
33
INTRODUCTION...............................................................................................................33
KINETICS OF SO
ASSEMBLY/DISASSEMBLY..............................................................................33
RESULTS..............................................................................................................................36
AN IS RECRUITED TO 80S BOTH ON APPLICATION AND REMOVAL OF
STRESS.................................36
AN REGULATES SGS DISSOLUTION VIA ITS NADH BINDING
DOMAIN.......................................38
ADDITION OF PYRUVATE CAN INDUCE CHANGES IN AN S
LOCALIZATION.....................................44
AN IS INVOLVED IN ABIOTIC STRESS RESPONSES IN
ARABIDOPSIS..............................................45
DISCUSSION......................................................................................................................
47
AN SENSES BOTH APPLICATION AND REMOVAL OF STRESS
..........................................................
47
AN IS A NOVEL SO DISASSEMBLING FACTOR IN
PLANTS...........................................................48
LINK BETWEEN STRESS, SGS AND NADH METABOLISM: HOW AN MIGHT REGULATE SO
DISASSEMBLY?.....................................................................................................................
49
AN IS INVOLVED IN SALT AND OSMOTIC STRESS RESPONSES IN ARABIDOPSIS
...............................
51
ENHANCER OF DECAPPING 3 (EDC3) AND AN: TWO STRUCTURALLY DIVERSE NADH
BINDING
PROTEINS INVOLVED IN THE REGULATION OF MRNP GRANULES
....................................................
51
CHAPTER 3. AIK, A NOVEL PROTEIN KINASE LOCALIZES TO
SGS.......................................................53
INTODUCTION...................................................................................................................53
RESULTS..............................................................................................................................54
AIK LOCALIZES TO MRNP
GRANULES......................................................................................54
AN AND AIK INTERACT WITH EACH OTHER IN LUMIER AND A MUTATION IN THE NADH
BINDING
POCKET DISRUPTS THIS
INTERACTION.........................................................................................56
AN AND AIK DO NOT AFFECT EACH OTHER S LOCALIZATION TO
SGS...........................................57
MOLECULAR ANALYSIS OF DIFFERENT DOMAINS OF AIK
.............................................................
59
COMPARISON OF THE INTERACTION BETWEEN AN AND TRUNCATIONS OF AIK
.............................
62
DISCUSSION......................................................................................................................
65
AIK, PLANT HOMOLOGUE OF DYRK FAMILY LOCALIZES TO SGS
.............................................
65
NADH BINDING DOMAIN IS CRUCIAL FOR INTERACTION BETWEEN AN AND AIK
........................
66
CHAPTER 4. MOLECULAR CHARACTERIZATION OF AN S NADH BINDING DOMAIN
..............................
68
INTRODUCTION
68
RESULTS.............................................................................................................................68
AN S DIMERIZATION IS COMPROMISED WITH A MUTATION IN NADH BINDING DOMAIN
...........
68
DIRECT ADDITION OF NADH DOES NOT CHANGE DIMERIZATION LEVELS OF AN
...........................
71
MAJORITY OF AN IS PROBABLY ALREADY IN A DIMERIZED STATE WHEN EXTRACTED
FROM HEK CELLS
...........................................................................................................................................73
DIRECT ADDITION OF NADH DOES NOT CHANGE THE DIMERIZATION OF N AND
C-TERMINUS
DOMAINS OF
AN..................................................................................................................73
COMPARISON OF DIMERIZATION EFFICIENCY OF DIFFERENT DOMAINS OF AN
..............................
74
PURIFICATION AND CHARACTERIZATION OF ANTI-AN
ANTIBODY....................................................75
DISCUSSION.......................................................................................................................78
DISRUPTION OF NADH BINDING POCKET INFLUENCES AN S DIMERIZATION
..............................
78
IS DIMERIZATION IMPORTANT FOR AN S
FUNCTION?...................................................................79
KEY FINDINGS AND
CONCLUSIONS...............................................................................80
A HYPOTHETICAL MODEL EXPLAINING AN S ROLE IN REGULATING SG KINETICS
...................................
82
MATERIALS AND METHODS
............................................................................................
83
MATERIALS........................................................................................................................83
CHEMICALS AND
ENZYMES....................................................................................................83
BACTERIAL
STRAINS.................................................................................................................83
PLANT
LINES..........................................................................................................................83
PRIMERS..............................................................................................................................83
BASIC VECTORS USED IN THE
STUDY........................................................................................84
LIST OF
CONSTRUCTS...............................................................................................................84
LIST OF TRANSGENIC
LINES......................................................................................................84
METHODS...........................................................................................................................85
TRANSFORMATION OF ARABIDOPSIS CELL
CULTURE.....................................................................85
STREP-TAG PURIFICATION OF AN FROM ARABIDOPSIS CELL
CULTURE.............................................85
PULLDOWN EXPERIMENT USING LUMIER
..............................................................................
85
LUMIER INVOLVING NADH
TREATMENTS............................................................................86
FUSION PCR FOR SITE-DIRECTED
MUTAGENESIS.......................................................................86
CONFOCAL IMAGING
SALT AND OSMOTIC STRESS TREATMENT OF
SEEDLINGS.................................................................87
PLANT
TRANSFORMATION...........................................................................................................87
TRANSIENT EXPRESSION
ASSAY...............................................................................................87
YEAST TWO
HYBRID................................................................................................................87
RECOMBINANT PROTEIN
PURIFICATION.....................................................................................87
GEL
FILTRATION.......................................................................................................................88
ANTIBODY
PURIFICATION........................................................................................................
88
PREPARATION OF PROTEIN EXTRACTS FROM THE PLANTS FOR WESTERN BLOT
.....................................
88
WESTERN BLOT
DETECTION......................................................................................................
89
PEPTIDE MASS
FINGERPRINTING.............................................................................................89
REFERENCES..........................................................................................................................90
APPENDIX........................................................................
97
ERKLAERUNG......................................................................................................................105
LEBENSLAUF.......................................................................................................................107
|
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| author | Bhasin, Hemal |
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| dewey-search | 572.452364 |
| dewey-sort | 3572.452364 |
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| discipline | Biologie |
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| spelling | Bhasin, Hemal Verfasser aut Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana vorgelegt von Hemal Bhasin Köln 2017 2 Mikrofiches (XIII, 107 Seiten) Illustrationen, Diagramme txt rdacontent h rdamedia he rdacarrier 24x Dissertation Universität zu Köln 2017 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1131737814/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029889708&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Bhasin, Hemal Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana |
| subject_GND | (DE-588)4113937-9 |
| title | Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana |
| title_auth | Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana |
| title_exact_search | Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana |
| title_full | Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana vorgelegt von Hemal Bhasin |
| title_fullStr | Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana vorgelegt von Hemal Bhasin |
| title_full_unstemmed | Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana vorgelegt von Hemal Bhasin |
| title_short | Molecular-functional characterization of Angustifolia gene and its role in regulating stress granule kinetics in Arabidopsis thaliana |
| title_sort | molecular functional characterization of angustifolia gene and its role in regulating stress granule kinetics in arabidopsis thaliana |
| topic_facet | Hochschulschrift |
| url | http://d-nb.info/1131737814/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029889708&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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