Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | Volltext Inhaltsverzeichnis |
| Item Description: | Zsfassung in dt. und engl. Sprache. - Online erschienen: 2014 |
| Physical Description: | 137 S. Ill., graph. Darst. |
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| 100 | 1 | |a Stegmann, Martin |d 1983- |e Verfasser |0 (DE-588)1049910257 |4 aut | |
| 245 | 1 | 0 | |a Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity |c von Martin Stegmann |
| 246 | 1 | 3 | |a Identifizierung von PUB22 Zielproteinen und deren funktionelle Charakterisierung in der PAMP-vermittelten Immunantwort |
| 264 | 1 | |c 2013 | |
| 300 | |a 137 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Zsfassung in dt. und engl. Sprache. - Online erschienen: 2014 | ||
| 502 | |a Würzburg, Univ., Diss., 2013 | ||
| 546 | |a Mit einer Zssfassung in dt. Sprache | ||
| 650 | 0 | 7 | |a Signaltransduktion |0 (DE-588)4318717-1 |2 gnd |9 rswk-swf |
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| 650 | 0 | 7 | |a Ubiquitin-Protein-Ligase |0 (DE-588)4398417-4 |2 gnd |9 rswk-swf |
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| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 912 | |a ebook | ||
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Record in the Search Index
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|---|---|
| adam_text | INDEX
INDEX
LIST OF ABBREVIATIONS 1
1. INTRODUCTION 1
1.1. PLANT IMMUNITY 1
1.2. VESICULAR TRAFFICKING IN PLANT IMMUNITY 6
1.3. THE PLANT EXOCYST COMPLEX 9
1.4. THE
UBIQUITIN MODIFICATION SYSTEM 11
1.4.1. PLANT U-BOX PROTEINS
(PUBS) 14
1.4.2. UBIQUITINATION IN PLANT IMMUNITY 16
1.4.3. NEGATIVE REGULATION
OF PTI BY PUB2Z, PUB23 AND PUB24 19
1.5. AIM OF THE PRESENT WORK 22
2. MATERIALS AND METHODS 23
2.1. MATERIALS 23
2.1.1. CHEMICALS 23
2.1.2. MEDIA 23
2.1.3. PLANTS 24
2.1.4. BACTERIA:. 24
2.1.5. HYALOPERONOSPORA ARABIDOPSIDIS 25
2.2. METHODS 25
2.2.1. CLONING 25
2.2.2. TRANSFORMATIONS 26
2.2.3. PLASMID PREPERATIONS 29
2.2.4. YEAST TWO-HYBRID SCREEN 29
2.2.5. YEAST TWO-HYBRID ANALYSIS AND YEAST COMPLEMENTATION
ASSAYS 29
2.2.6. BIMOLECULAR FLUORESCENCE COMPLEMENTATION 30
2.2.7. PROTEIN ISOLATION
FROM PLANTS 30
2.2.8. COIMMUNOPRECIPITATION 31
2.2.9. IN VITRO ASSAYS
WITH RECOMBINANT
PROTEINS 32
2.2.10. QUANTITATIVE REAL-TIME
PCR 34
2.2.11. GENOTYPING
OF T-DNA INSERTION
MUTANTS 34
2.2.12. ROS-BURST ASSAYS 35
2.2.13. MAPK ASSAYS 36
2.2.14. ANALYSIS OF PAMP-INDUCED
GENE EXPRESSION 36
2.2.15. ROOT
GROWTH INHIBITION ASSAYS 36
2.2.16. PATHOGEN INFECTION
ASSAYS 37
2.2.17. STATISTICAL DATA
ANALYSIS 38
HTTP://D-NB.INFO/1049910303
3. RESULTS 39
3.1. IDENTIFICATION OF PUB22 CANDIDATE TARGETS 39
3.1.1. YEAST TWO-HYBRID SCREEN TO IDENTIFY CANDIDATE PUB22 INTERACTORS
39
3.1.2. INTERACTION ANALYSIS OF PUB22 WITH ADDITIONAL SUBUNITS OF THE
EXOCYST COMPLEX BY YEAST TWO-
HYBRID ASSAY 43
3.1.3. YEAST ASSAY TO TEST THE ABILITY OF EXO70B2 TO COMPLEMENT YEAST
EXO70
DEFICIENCY 44
3.2. ANALYSIS OF THE IMPACT OF PUB22 CANDIDATE TARGETS ON PLANT IMMUNITY
45
3.2.1. GENERATION
OF T-DNA INSERTION
MUTANTS OF CANDIDATE PUB22 TARGETS 45
3.2.2. ROS-BURST ASSAYS
OF CANDIDATE PUB22 INTERACTOR
MUTANTS 46
3.2.3. PATHOGEN GROWTH
ASSAYS WITH CANDIDATE
PUB22 INTERACTOR MUTANTS 47
3.2.4. GENE EXPRESSION
PROFILE UPON ELICITOR TREATMENT OF CANDIDATE
PUB22 INTERACTORS SO
3.3. CONFIRMATION OF THE INTERACTION BETWEEN PUB22 AND EXO70B2 51
3.3.1. CONFIRMATION OF THE INTERACTION OF PUB22 AND EXO70B2 BY
BIMOLECULAR FLUORESCENCE
COMPLEMENTATION 51
3.3.2. CONFIRMATION
OF THE PUB22-EXO70B2 INTERACTION BY COIMMUNOPRECIPITATION 54
3.3.3. ANALYSIS OF A PHYSICAL
INTERACTION OF PUB22
AND EXO70B2 BY IN VITRO PULL-DOWN 55
3.3.4. INTERACTION ANALYSIS OF EXO70B2 WITH PUB23 AND PUB24 USING
BIMOLECULAR FLUORESCENCE
COMPLEMENTATION 56
3.4. ANALYSIS OF FLG22-DEPENDENT PUB22 PROTEIN STABILIZATION 58
3.4.1. HA-PUB22 PROTEIN ACCUMULATION AFTER TRANSIENT OVEREXPRESSION IN
NICOTIANA BENTHAMIANA AND
ARABIDOPSIS THALIANA MESOPHYLL PROTOPLASTS 58
3.4.2. TIME-COURSE ANALYSIS OF HA-PUB22 PROTEIN ACCUMULATION AFTER FLG22
ELICITATION OR PROTEASOME
INHIBITION 60
3.5. ANALYSIS OF THE UBIQUITINATION AND DEGRADATION OF EXO70B2 BY PUB22
63
3.5.1. IN VITRO
UBIQUITINATION OF EXO70B2
BY PUB22 63
3.5.2. ANALYSIS OF PUB22-MEDIATED EXO70B2 DEGRADATION BY BIMOLECULAR
FLUORESCENCE
COMPLEMENTATION 64
3.5.3. EXPRESSION OF CMYC-EXO70B2 IN WILD TYPE AND PUB22/23/24
ARABIDOPSIS THALIANA MESOPHYLL
PROTOPLASTS 67
3.6. CHARACTERIZATION OF THE PAMP-TRIGGERED RESPONSES AND DISEASE
RESISTANCE OF EXO7062
MUTANTS 69
3.6.1. ISOLATION OF TWO INDEPENDENT EXO70B2 T-DNA INSERTION LINES AND
GENERATION OF A
PUB22/23/24/EXO70B2 QUADRUPLE MUTANT 69
3.6.2. ANALYSIS OF PAMP-TRIGGERED RESPONSES OF EXO70B2 MUTANTS 71
3.6.3. DISEASE RESISTANCE
ANALYSIS OF EXO70B2 MUTANTS 78
3.7. FUNCTIONAL REDUNDANCY ANALYSIS OF THE EXO70B2 HOMOLOG EXO70B1 83
3.7.1. GENERATION OF TWO INDEPENDENT EXO70B1 T-DNA INSERTION LINES AND
AN EXO70B1/EXO70B2
DOUBLE MUTANT 83
3.7.2. ANALYSIS OF PAMP-TRIGGERED RESPONSES OF EXO70B1 MUTANTS 85
3.7.3. DISEASE
RESISTANCE ANALYSIS OF EXO70B1
MUTANTS 87
DISCUSSION.
4.1. PUB22 TARGETS THE VESICULAR TRAFFICKING COMPONENT EXO70B2 TO
DOWNREGULATE PTI SIGNALING
89
4.2. ADDITIONAL POTENTIAL TARGET PROTEINS OF PUB22 SUGGEST A
SPECIALIZATION IN THE REGULATION OF
VESICULAR TRAFFICKING 99
4.3. ADDITIONAL DEFENSE RESPONSE PATHWAYS POTENTIALLY TARGETED BY PUB22
103
4.4. THE FUNCTION OF PUB22 IS REGULATED BY POSTTRANSLATIONAL PROTEIN
STABILIZATION 104
4.5. WORKING MODEL FOR THE FUNCTION OF PUB22 AND EXO70B2 IN REGULATING
PTI RESPONSES 108
5. SUMMARY 110
6. ZUSAMMENFASSUNG 111
7. BIBLIOGRAPHY 112
8. APPENDIX 130
|
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| discipline | Biologie |
| format | Thesis Book |
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| spelling | Stegmann, Martin 1983- Verfasser (DE-588)1049910257 aut Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity von Martin Stegmann Identifizierung von PUB22 Zielproteinen und deren funktionelle Charakterisierung in der PAMP-vermittelten Immunantwort 2013 137 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache. - Online erschienen: 2014 Würzburg, Univ., Diss., 2013 Mit einer Zssfassung in dt. Sprache Signaltransduktion (DE-588)4318717-1 gnd rswk-swf Exocytose (DE-588)4276172-4 gnd rswk-swf Ubiquitin-Protein-Ligase (DE-588)4398417-4 gnd rswk-swf Yeast-Two-Hybrid-System (DE-588)4554290-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Exocytose (DE-588)4276172-4 s Ubiquitin-Protein-Ligase (DE-588)4398417-4 s Yeast-Two-Hybrid-System (DE-588)4554290-9 s Signaltransduktion (DE-588)4318717-1 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-92061 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-92061 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027252771&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Stegmann, Martin 1983- Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity Signaltransduktion (DE-588)4318717-1 gnd Exocytose (DE-588)4276172-4 gnd Ubiquitin-Protein-Ligase (DE-588)4398417-4 gnd Yeast-Two-Hybrid-System (DE-588)4554290-9 gnd |
| subject_GND | (DE-588)4318717-1 (DE-588)4276172-4 (DE-588)4398417-4 (DE-588)4554290-9 (DE-588)4113937-9 |
| title | Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity |
| title_alt | Identifizierung von PUB22 Zielproteinen und deren funktionelle Charakterisierung in der PAMP-vermittelten Immunantwort |
| title_auth | Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity |
| title_exact_search | Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity |
| title_full | Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity von Martin Stegmann |
| title_fullStr | Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity von Martin Stegmann |
| title_full_unstemmed | Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity von Martin Stegmann |
| title_short | Identification of PUB22 Targets and Functional Characterization in PAMP-Triggered Immunity |
| title_sort | identification of pub22 targets and functional characterization in pamp triggered immunity |
| topic | Signaltransduktion (DE-588)4318717-1 gnd Exocytose (DE-588)4276172-4 gnd Ubiquitin-Protein-Ligase (DE-588)4398417-4 gnd Yeast-Two-Hybrid-System (DE-588)4554290-9 gnd |
| topic_facet | Signaltransduktion Exocytose Ubiquitin-Protein-Ligase Yeast-Two-Hybrid-System Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-92061 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027252771&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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