Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana:
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| Format: | Thesis Book |
| Language: | English German |
| Published: |
2013
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Zsfassung in dt. Sprache |
| Physical Description: | IX, 134 S. Ill., graph. Darst. |
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MARC
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| 100 | 1 | |a Lee, Hye-Jung |d 1979- |e Verfasser |0 (DE-588)103414488X |4 aut | |
| 245 | 1 | 0 | |a Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana |c von Hye-Jung Lee |
| 264 | 1 | |c 2013 | |
| 300 | |a IX, 134 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
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| 500 | |a Zsfassung in dt. Sprache | ||
| 502 | |a Berlin, Humboldt-Univ., Diss., 2013 | ||
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| adam_text | IMAGE 1
TABLE O F C O N T E N T S
T A B L E OF CONTENTS
T A B L E O F C O N T E N T S I
A B B R E V I A T I O N S V
Z U S A M M E N F A S S U N G V I I I
A B S T R A C T I X
1. I N T R O D U C T I O N 1
I. I H E M E IN HIGHER PLANTS 2
1.1.1 HEME SYNTHESIS 2
1.1.2 H E M E DEGRADATION 4
1.2 REGULATORY ROLES O F H E M E IN CELLULAR METABOLISMS 7
1.2.1 REGULATION IN A L A FORMATION 7
1.2.2 RETROGRADE SIGNALING PATHWAY 8
1.3 FUNCTIONAL SIGNIFICANCE O F H E M E O X Y G E N A S E 1 IN PLANTS 9
1.3.1 SYNTHESIS O F PHYTOCHROME PRECURSOR 10
1.3.2 H E M E O X Y G E N A S E 1 IS INVOLVED IN MOLECULAR ACTION UNDER
ENVIRONMENTAL CHANGES 10
1.3.3 PLANT R O O T D E V E L O P M E N T 12
1.4 IDENTIFICATION O F N E W MEMBERS O F CYTOSOLIC H E M E - B I N D I
N G PROTEINS IN VERTEBRATES 13
1.4.1 STRUCTURAL ASPECTS O F P 2 2 H B P AND S O U L PROTEINS 14
1.4.2 FUNCTIONS O F M E M B E R S O F S O U L / H E M E - B I N D I N G
PROTEIN FAMILY IN M A M M A L S 16
1.5 G O A L O F THIS STUDY 19
2. M A T E R I A L S A N D M E T H O D S 2 1
2.1 CHEMICALS, KITS A N D E N Z Y M E S 21 ..
2.2 E Q U I P M E N T 21
2.3 PLANT MATERIAL AND GROWTH CONDITIONS 2 2
2.4 MEDIA, SOLUTIONS AND BUFFERS 2 2
2.5 STANDARD PROTOCOLS F O R MOLECULAR BIOLOGY 2 5
2.5.1 POLYMERASE CHAIN REACTION 2 5
2.5.2 PREPARATION O F CHEMICALLY COMPETENT E. COLI CELLS 2 5
2.5.3 TRANSFORMATION O F CHEMICALLY COMPETENT E. COLI CELLS 2 6 ,
2.5.4 COLONY P C R 2 7
2.5.5 ISOLATION O F PLASMIDS FROM /** . COLI CELLS 2 7
2.5.6 PREPARATION O F CHEMICALLY COMPETENT AGROBACTERIUM TUMEFACIENS
CELLS 2 8
2.5.7 TRANSFORMATION F O R CHEMICALLY COMPETENT A G R O B A C T E R I U
M TUMEFACIENS 2 8
I
HTTP://D-NB.INFO/1036577120
IMAGE 2
TABLE O F C O N T E N T S
2 . 5 . 8 S T A B L E TRANSFORMATION O F ARABIDOPSIS U S I N G
FLOWER-DIP M E T H O D 2 9
2.5.9 ISOLATION O F G E N O M I C D N A F R O M ARABIDOPSIS PLANTS 2 9
2 5 10 ISOLATION O F R N A FROM ARABIDOPSIS PLANTS AND GEL
ELECTROPHORESIS 2 9
2 5 11 M E A S U R E M E N T O F N U C L E O T I D E S C O N C E N T R A
T I O N 3 0
2.5.12 SYNTHESIS O F C D N A FROM R N A 3 0
2.5.13 EXTRACTION O F TOTAL PROTEINS FROM ARABIDOPSIS PLANTS 31
2 5.14 D E T E R M I N A T I O N O F P R O T E I N S C O N C E N T R A T
I O N BY T H E BRADFORD M E T H O D 31
2.5.15 W E S T E R N B L O T A N A L Y S I S 31
2 . 5 . 1 6 COLLECTION O F T - D N A MUTANTS 3 2
2.5.17 G E N O T Y P I N G O F T - D N A M U T A N T S 3 3
2.5.18 PRIMERS SYNTHESIS AND D N A S E Q U E N C I N G 3 3
2.6 R E C O M B I N A N T PROTEIN EXPRESSION IN E. COLI CELLS 3 3
2.6.1 CLONING O F A T H B P GENES AND CONSTRUCTIONS O F EXPRESSION
VECTORS 3 3
2.6.2 EXPRESSION O F RECOMBINANT A T H B P S IN E. COLI CELLS 3 4
2.6.3 EXPRESSION A N D PURIFICATION O F RECOMBINANT PROTEINS FROM E.
COLI CELLS U N D E R NATIVE
CONDITIONS 3 5
2.6.4 PREPARATION O F RECOMBINANT A T H B P 5 PROTEIN F O R ANTIBODY
PRODUCTION 3 5
2.7 H E M E - B I N D I N G ASSAYS O F A T H B P S 3 6
2.7.1 I N VITRO BINDING ASSAY USING HEMIN-AGAROSE 3 6
2.7.2 UV-VISIBLE SPECTRA 3 6
2.8 PRODUCTION O F ARABIDOPSIS TRANSFORMANTS CARRYING FUSION G U S
CONSTRUCTS AND G U S
HISTOCHEMICAL STAINING 3 7
2.8.1 PREPARATION O F 5 - U T R ATHBP::G )S REPORTER CONSTRUCTS 3 7
2.8.2 PRODUCTION AND SELECTION O F G U S - A R A B I D O P S I S
TRANSFORMANTS 3 8
2.8.3 G U S HISTOCHEMICAL STAINING 3 8
2.9 PROTEIN SUBCELLULAR LOCALIZATION USING FLUORESCENCE M A R K E R 3 9
2.9.1 PREPARATION O F RED FLUORESCENT PROTEIN FUSION CONSTRUCTS 3 9
2.9.2 GENERATION O F BINARY VECTORS 4 0
2.9.3 MICROSCOPIC ANALYSIS 4 0
2.10 PRODUCTION OF/FRA/BP5-OVEREXPRESSING TRANSGENIC ARABIDOPSIS PLANTS
4 1
2.11 YEAST TWO-HYBRID SYSTEM 4 2
2.11.1 CONSTRUCTION O F A T H B P S A N D A T H Y L G E N E S IN BAIT A
N D PREY VECTORS 4 2
2.11.2 TRANSFORMATION O F Y E A S T CELLS 4 3
2.11.3 CO-TRANSFORMATION 4 3
2.11.4 YEAST TWO-HYBRID ASSAY AFTER CO-TRANSFORMANT 4 3
2.12 BIMOLECULAR FLUORESCENCE C O M P L E M E N T A T I O N 4 4
II
IMAGE 3
TABLE O F C O N T E N T S
2.12.1 CONSTRUCTIONS O F RECOMBINANT BINARY VECTORS 4 4
2.12.2 TRANSIENT EXPRESSION IN TOBACCO LEAVES BY AGROBACTERIUM
INFILTRATION 4 4
2.12.3 MICROSCOPIC ANALYSIS 4 5
2.13 M E A S U R E M E N T O F H E M E CONTENT IN PLANTS 4 5
2.14 DETECTION O F HYDROGEN PEROXIDE 4 6
2.14.1 D A B HISTOCHEMICAL STAINING WITH INTACT PLANTS 4 6
2.14.2 M E A S U R E M E N T O F HYDROGEN PEROXIDE CONCENTRATION IN
PLANTS 4 6
2.15 SEMI-QUANTITATIVE R T - P C R 4 7
2.16 REAL-TIME P C R FOR RELATIVE EXPRESSION O F GENES 4 7
3. R E S U L T S 4 9
3.1 A FAMILY O F P22HBP/SOUL-RELATED PROTEIN IN A R A B I D O P S I S
THALIANA 4 9
3.1.1 ARABIDOPSIS PROTEINS WITH A CONSERVED SOUL/HEME-BINDING PROTEIN D
O M A I N 4 9
3.1.2 IN SILICO LOCATION O F T H E H B P GENES IN THE A R A B I D O P S
I S THALIANA G E N O M E 51
3.1.3 ANALYSIS O F T H E SECONDARY STRUCTURE O F THE ARABIDOPSIS H B P
PROTEINS 52
3.1.4 THE PHYLOGENETIC TREE O F P 2 2 H B P / S O U L - R E L A T E D
PROTEINS IN VASCULAR PLANTS 5 3
3.2 MOLECULAR INTERACTION BETWEEN M E M B E R S O F ARABIDOPSIS H B P
FAMILY A N D H E M E 5 6
3.2.1 T H R E E ARABIDOPSIS H B P S H A V E BINDING AFFINITY TO
HEMIN-AGAROSE 5 6
3.2.2 VERIFICATION O F HEME-BINDING AFFINITY O F ATL 1 HPS 5 8
3.3 TISSUE A N D CELLULAR DISTRIBUTION O F ARABIDOPSIS H B P FAMILY M E
M B E R S 5 9
3.3.1 T H E M E M B E R S O F ARABIDOPSIS H B P G E N E FAMILY S H O W
DISTINCT AND SPECIFIC EXPRESSION
PATTERNS D U R I N G P L A N T DEVELOPMENT 5 9
3.3.2 DISTINCT SUBCELLULAR LOCALIZATION O F M E M B E R S O F
ARABIDOPSIS H B P FAMILY 6 2
3.3.3 A T H B P 3 AND A T L L B P S ARE CHLOROPLAST-TARGETED PROTEINS 6
3
3.4 TRANSCRIPTIONAL REGULATIONS O F ARABIDOPSIS H B P FAMILY IN RESPONSE
TO EXOGENOUS AGENTS 6 5
3.4.1 H E M I N UPTAKE FROM THE GROWTH M E D I U M 6 6
3.4.2 INDUCIBLE EXPRESSION O F A T H B P L FOLLOWING H E M I N TREATMENT
6 6
3.4.3 A T H B P L EXPRESSION IS AFFECTED BY TREATMENT WITH H E M I N AND
I L O ; 6 8
3.4.4 A T H B P 3 EXPRESSION IS INDUCED BY H 2 0 2 B U T REPRESSED BY H
E M I N TREATMENT 6 9
3.4.5 EXPRESSION O F A T H B P 5 IS UNAFFECTED B Y H E M I N FEEDING 7 0
3.4.6 A T H B P 5 A N D A T H Y L TRANSCRIPTS RESPOND SIMILARLY T O
OXIDATIVE STRESS 7 2
3.5 CHARACTERIZATION O F THE BIOLOGICAL ROLE O F A T H B P 5 IN H E M E
CATABOLISM 7 3
3.5.1 IDENTIFICATION O F AN A T H B P 5 INTERACTING PROTEIN BY YEAST
TWO-HYBRID ANALYSIS 7 3
3.5.2 VERIFICATION O F INTERACTION BETWEEN A T H B P 5 AND A T H Y L I N
P L A N T A 7 5
3.5.3 ISOLATION O F A H O M O Z Y G O U S A T H B P S MUTANT 7 7
3.5.4 MOLECULAR ANALYSES WITH THE ATHBP5 MUTANT LINE 7 8
3.5.5 T H E A T H B P 5 MUTANT IS SUSCEPTIBLE T O OXIDATIVE STRESS 7 9
III
IMAGE 4
TABLE O F C O N T E N T S
3 . 5 . 6 ANALYSIS OF.!(///F/ .5-OVERCXPRCSSING ARABIDOPSIS PLANTS 81
3 5 7 T H E A T H B P S OVER-EXPRESSING PLANTS ARE RESISTANT T O
OXIDATIVE STRESS 8 3
3 5.8 H E M E CONTENT W A S INCREASED IN T H E A T H B P 5 MUTANT 8 4
4. D I S C U S S I O N 8 5
4 I I D E N T I F I C A T I O N O F A N O V E L SIX M E M B E R H B P G
E N E F A M I L Y I N T H E / ) , THALIANA G E N O M E 8 5
4 2 T H E SECONDARY FOLDING PATTERN O F T H E M A M M A L I A N P 2 2 H
B P PROVIDES A MODEL F O R BINDING O F T H E
ARABIDOPSIS H B P S TO CYCLIC TETRAPYRROLES 8 6
4.3 PHYLOGENETIC RELATIONSHIP O F H B P S F R O M H I G H E R PLANTS
INDICATES THAT ARABIDOPSIS H B P S CLUSTER
INTO T H E FOUR CLADES 8 7
4.4 T H R E E MEMBERS O F T H E ARABIDOPSIS H B P FAMILY BIND H E M E IN
VITRO 8 8
4.5 PROMOTER-GUS ANALYSIS REVEALS REGULATION O F A T H B P S GENE
EXPRESSION DURING PLANT GROWTH A N D
DEVELOPMENT 8 9
4 . 6 A T H B P L M A Y PLAY A ROLE IN BINDING CYCLIC TETRAPYRROLES AT
THE VACUOLAR M E M B R A N E 91
4.7 RAPID AND D Y N A M I C REGULATION O F A T H B P 2 IS REQUIRED T O
PROMOTE CELLULAR RESPONSES T O
ENVIRONMENTAL CHANGES 9 3
4.8 A T H B P 3 M A Y FUNCTION IN PLASTOGLOBULES ASSOCIATED WITH PLANT A
G I N G A N D STRESS 9 4
4.9 A T H B P 5 PROTEIN M A Y BE INVOLVED IN H E M E DISTRIBUTION IN
CHLOROPLASTS 9 8
4.9.1 T H E PHENOTYPE O F A T H B P S DOES N O T COPY H Y L 9 8
4.9.2 MANIPULATION O F THE A T H B P S G E N E AFFECTS A CELLULAR
DEFENSE RESPONSE T O R O S 9 9
4.10 REGULATION O F H E M E CATABOLISM IN PRESENCE O F EXCESS H E M E
AND H2O2 101
4.10.1 TRANSCRIPTIONAL REGULATION O F A T H Y L M A Y BE DEPENDENT O N
SIGNALS FROM DIFFERENT H E M E
POOLS 102
4.10.2 A T H B P 5 AND A T H Y L A R E INDUCED IN RESPONSE TO R O S 103
5. R E F E R E N C E S 106
S U P P L E M E N T A R Y T A B L E S I 123
S U P P L E M E N T A R Y T A B L E S2 126
S U P P L E M E N T A R Y T A B L E S3 127
S U P P L E M E N T A R Y FIG. SI 128
S U P P L E M E N T A R Y FIG. S2 129
A C K N O W L E D G E M E N T S 130
S E L B S T A N D I G K E I T S E R K L A R U N G 131
C U R R I C U L U M V I T A E 132
IV
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| author_facet | Lee, Hye-Jung 1979- |
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| author_sort | Lee, Hye-Jung 1979- |
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| dewey-ones | 572 - Biochemistry |
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| indexdate | 2024-07-10T00:36:36Z |
| institution | BVB |
| language | English German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-025953580 |
| oclc_num | 844088142 |
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| physical | IX, 134 S. Ill., graph. Darst. |
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| spelling | Lee, Hye-Jung 1979- Verfasser (DE-588)103414488X aut Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana von Hye-Jung Lee 2013 IX, 134 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. Sprache Berlin, Humboldt-Univ., Diss., 2013 (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=025953580&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lee, Hye-Jung 1979- Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana |
| subject_GND | (DE-588)4113937-9 |
| title | Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana |
| title_auth | Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana |
| title_exact_search | Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana |
| title_full | Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana von Hye-Jung Lee |
| title_fullStr | Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana von Hye-Jung Lee |
| title_full_unstemmed | Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana von Hye-Jung Lee |
| title_short | Molecular and functional characterization of the heme-binding protein (HBP) family in Arabidopsis thaliana |
| title_sort | molecular and functional characterization of the heme binding protein hbp family in arabidopsis thaliana |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025953580&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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