Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1:
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| Format: | Thesis Book |
| Language: | English |
| Published: |
Berlin
2015
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | X, 144 Seiten Illustrationen, Diagramme |
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| 245 | 1 | 0 | |a Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 |c vorgelegt von: Sonja Maria Sachse aus Berlin |
| 264 | 1 | |a Berlin |c 2015 | |
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| 502 | |b Dissertation |c Freie Universität Berlin |d 2015 | ||
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| adam_text | CONTENTS
1.PART1 LUTERATUREREVIEW 1
1.1 INTRODURT*ON 2
1.2 DSCAMS ARE NEURONA* CE** ADHESION MOLECULES OF THE IMMUNOGLOBULIN
SUPERFAMILY 5
1.3 MOLECULAR DIVERSITY IS ESSENTIA, FOR NEURAL CIRCUIT FORMATION 6
1 *3.1 THE DROSOPHILA DSCAML LOCUS GENERATES EXTRAORDINARY MOLECULAR
DIVERSITY 6
1.3.2 DROSOPHILA DSCAML ISOFORMS EXHIBIT EXTRAORDINARY HOMOPHILIC
BINDING SPECIFICITIES 6
1.3.3 ISOFORM-SPECJFIC HOMOPHILJC BINDING OF DSCAML PROVIDES THE
MOLECULAR BASIS FOR SELF-AVOIDANCE 8
1 *3.4 DIVERSITY OF DSCAML IS ESSENTIAL FOR NEURITE SELF-VERSUS
NONSELF-DISCRIMINATION 10
1.3.5 VERTEBRATE DSCAM GENES DO NOT ENCODE A VAST AMOUNT OF ISOFORMS 11
1.3.6 CLUSTERED PROTOCADHERINS ESTABLISH MOLECULAR DIVERSITY IN THE
VERTEBRATE NERVOUS SYSTEM 11
1.4 VERTEBRATE DSCAMS ARE EXPRESSED IN THE DEVELOPING AND ADULT NERVOUS
SYSTEM 13
1.5 DSCAMS PROMOTE NEURONAL SELF-AVOIDANCE AND TILING IN THE VERTEBRATE
RETINA 14
1
*
6 DSCAMS PLAY A ROLE *N NEURITE BRANCHING 15
1.7 DSCAM ACTS AS A RECEPTOR SO
*
AXON GUIDANCE CUES 16
1.7.1 DSCAM IS A NETRIN RECEPTOR AND COLLABORATES WITH UNC5 AND DCC 16
1
*
8 DSCAMS PLAY A ROLE *N SYNAPTOGENESIS 18
1.9 DSCAMS PROMOTE PROGRAMMED CELL DEATH 20
1.9.1 PROGRAMMED CELL DEATH *S ESSENTIAL FOR PROPER NEURONAL WIRING 20
1.9.2 DSCAMS DRIVE NEURAL CELL DEATH IN THE DEVELOPING MOUSE RETINA 20
1.10 DSCAM
IS
ASSOCIATED WITH THE INTELLECTUAL DISABILITY AND CARDIAC DESERTS IN DOWN
SYNDROME 22
1.11 SIGNALING DOWNSTREAM OF DSCAM FAMILY *G-SF CAMS 24
1.11.1 SIGNALING DOWNSTREAM OF DROSO* 24
1.11.2 SIGNALING DOWNSTREAM OF VERTEBRATE DSCAMS 25
2 PART2
I
MATERIALAND METHODS 29
2.1 SEQUENCE INFORMATION AND SEQUENCE ALIGNMENTS 30
2.2 M OTIF PREDICTIONS 30
2.3 PLASMIDS AND MOLECULAR CLONING 31
2.3.1 POLYMERASE CHA*N REACTION AND SEQUENCING 31
2.3.2 DSCAM AND DSCAML1 CDNAS 31
2.3.3 GENERATION OF TAGGED DSCAM AND DSCAML1 FULL-LENGTH AND ICD
CONSTRUCTS 31
2.3.4 GENERATION OF THE MAPPIT PREY COLLECTION 32
2.3.5 GENERATION OF INDIVIDUAL MAPPIT BAITS AND PREYS 32
2.3.6 GENERATION OF MAPPIT BAIT AND PREY MUTANTS 32
2.3.7 GENERATION OF CLEAVAGE REPORTER ASSAY CONSTRUCTS 34
2.3.8 GENERATION OF RPTP CONSTRUCTS 34
2.4 MAPPIT 36
2.5 CELL CULTURE 36
2.5.1 VERTEBRATE CELL LINES 36
2.5.2 PRIMARY CULTURE OF MOUSE CORTICAL NEURONS 37
2.5.3 DROSOPHILA CELL CULTURE 37
2.6 TRANSFECTIONS 38
2.6.1 NUCLEOFECTION 38
2.6.2 GENERATION OF STABLE CELL LINES 38
2.7 IMMUNOPRECIPITATIONS AND IMMUNOBLOTTING 38
2.8 CLEAVAGE REPORTER ASSAY 39
2.9 DROSOPHILA TISSUE PROTEIN LYSATES 39
2.10 QUANTIFICATION OF DSCAML TYROSINE-PH.SPHORYLA.ION 40
2.10.1 GENERATION OF SHT CONDITIONED CELL CULTURE SUPERNATANT 40
2.10.2 QUANTIFICATION OF DSCAM TYROSINE-PHOSPHORYLATION 40
2.11 IMMUNOHISTOCHEMISTRY 41
2.12 IMAGE ACQUISITION AND ANALYSIS 41
2.13 STATISTICAL ANALYSIS 42
2.14 ISOLATION OF TOTAL RNA FOR MRNA SEQUENCING 42
2.15 MRNA SEQUENCING 42
2.15.1 RNA QUALITY CONTROL 42
2.15.2 LIBRARY PREPARATION 43
2.15.3 SEQUENCING 43
2.16 STATISTICAL ANALYSIS OF MRNA SEQUENCING DATA 43
2.16.1 PREPROCESSING 43
2.16.2 MAPPING 44
2.16.3 COUNTING 44
2.16.4 IDENTIFYING DIFFERENTIAL EXPRESSION 44
2.17 INGENUITY PATHWAY ANALYSIS 45
2.17.1 IPA CORE ANALYSIS 45
2.17.2 DOWNSTREAM EFFECTS ANALYSIS 45
2.17.3 UPSTREAM REGULATOR ANALYSIS 46
2.18 BUFFERS AND BACTERIAL MEDIA 46
3.PA R T3I RESULTS 49
3.1 VERTEBRATE DSCAMS EXHIBIT A VARIETY OF SIGNALING MOTIFS 50
3.1.1 DSCAM FAMILY MEMBERS EXHIBIT CONERVED SIGNALING MOTIFS 50
3.1.2 DSCAMS EXHIBIT DOCKING MOTIFS AND GENERAL UGAND BINDING SITES FOR
MODULAR PROTEIN DOMAINS 51
3.1.3 DSCAMS POSSESS SEVERAL POSTTRANSLATIONAL MODIFICATION SITES AND
DEGRON MOTIFS 51
3.1.4 DSCAMS EXHIBIT SUB-CELLULAR TARGETING AND CLEAVAGE SITES 54
3.1.5 DSCAMS EXHIBIT MULTIPLE TYROSINE KINASE SUBSTRATE SITES 55
3.2 A MAPPIT SCREEN REVEALS NOVEL SIGNALING TARGETS OF VERTEBRATE DSCAMS
57
3.2.1 A MAPPIT SCREEN IDENTIFIES DSCAM AND DSCAML1 INTERACTING PROTEINS
57
3.2.2 MUTATION OF TYROSINE-MOTIFS ABOLISHES THE INTERACTION WITH STAT3,
USP21 AND SH2D2A 59
3.2.3 DSCAMS INTERACT WITH THE DUAL-SPECIFICITY TYROSINE-REGULATED
KINASES DYRKI A AND DYRKI B 65
3.3 DSCAMS ARE CLEAVED AND THEIR INTRACELLULAR DOMAINS TRANSLOCATE INTO
THE NUCLEUS 66
3.3.1 A NUCLEAR LOCALIZATION SIGNAL MEDIATES THE INTERACTION BETWEEN
DSCAMS AND **05 66
3.3.2 THE INTRACELLULAR DOMAINS OF DSCAMS CAN TRANSLOCATE INTO THE
NUCLEUS 68
3.3.3 CLEAVAGE OF DSCAMS RESULTS IN THE LJBERATION OF INTRACEHULAR
DOMAIN FRAGMENTS 70
3.3.4 THE STABILITY OF DSCAMS MAY BE REGULATED BY UBIQUITIN-DEPENDED
PROTEASOMAL DEGRADATION 73
3.4 THE DSCAML1 ICD MODULATES THE EXPRESSION OF GENES INVOLVED *N NEURAL
CIRCUIT FORMATION
AND CELL DEATH 75
3.4.1 THE DSCAML1 ICD REGULATES THE TRANSCRIPTION OF GENES INVOLVED IN
AXON GUIDANCE SIGNALING 78
3.4.2 BDNF ACTS AS POTENTIAL UPSTREAM REGULATOR OF DSCAML1 INDUCED
TRANSCRIPTIONAL CHANGES 78
3.4.3 THE DSCAML1 ICD
S
INVOLVED IN THE TRANSCRIPTIONAL REGULATION OF NEURONAL CELL DEATH 80
3.5 OVEREXPRESSION OF THE DSCAML1 ICD LEADS TO IMPAIRED NEURITE
OUTGROWTH *N MOUSE CORT*CA* NEURONS 84
3.6 DSCAMS INTERACT WITH RECEPT.
*
TYROSINE PHOSPHATASES 86
3.6.1 SLIT DIRECTLY PROMOTES RPTP69D-DEPENDENT DSCAML DE-PHOSPHORYLATION
86
3.6.2 VERTEBRATE DSCAMS INTERACT WITH PTPRG AND PTPRE 88
4. PART4I DISCUSSION 91
4.1 A NOVE* M EM BRANE.TO-NUDEUS SIGNALING MECHANISM OF DSCAMS 92
4.1.1 THE DSCAML1 ICD MODULATES GENE TRANSCRIPTION 94
4.1.2 A POTENTIAL ROLE OF DSCAML1 IN TRANSCRIPTIONAL REGULATION OF
NEURONAL CELL DEATH? 95
4.1.3 DSCAML1 MAY REGULATE AXON GUIDANCE SIGNA^^ 98
4.1.4 POTENTIAL LIMITATIONS OF THE TRANSCRIPTOME ANALYSIS 99
4.1.5 A ROLE OF THE DSCAML1 ICD IN THE REGULATION OF NEURITE OUTGROWTH?
100
4.2 DSCAMS INTERACT WITH SERINEIIHREONINE KINASES AND SH2 ADAPTOR
PROTEINS 101
4.2.1 DSCAMS INTERACT WITH DYRK FAMILY SERINE/THREONINE KINASES 101
4.2.2 DSCAMS INTERACT WITH THE ADAPTOR PROTEIN SH2D2A 103
4.3 DSCAMS INTERACT WITH TRANSCRIPTIONALLY ACTIVE PARTNERS 104
4 .4 SIGNALING OF DSCAM FAMILY IGSF.CAMS IS REGULATED BY
POST-TRANSLATIONAL MODIFICATIONS 106
4.4.1 TYROSINE-PHOSPHORYLATION PLAYS A ROLE *N DSCAM SIGNALING 106
4.4.2 SIGNALING OF DSCAMS MIGHT BE REGULATED BY UBIQUITIN-MEDIATED
PROTEASOMAL DEGRADATION 108
4.5 CONCLUSIONS AND PERSPECTIVES 110
REFERENCES 113
ACKNOWLEDGMENTS 125
APPENDICES 127
APPENDIX 1 I SUPPLEMENTARY FIGURES 129
APPENDIX 2 I SUPPLEMENTARY INFORMATION 131
APPENDIX 3 I PUBLICATIONS AND PRESENTATIONS 138
APPENDIX 4 I CONTRIBUTIONS TO THE EXPERIMENTS 140
APPENDIX 5 I FUNDING 141
APPENDIX 6 I LIST OF ABBREVIATIONS 142
|
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| spelling | Sachse, Sonja Maria Verfasser (DE-588)1080911901 aut Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 vorgelegt von: Sonja Maria Sachse aus Berlin Berlin 2015 X, 144 Seiten Illustrationen, Diagramme txt rdacontent n rdamedia nc rdacarrier Dissertation Freie Universität Berlin 2015 Wirbeltiere (DE-588)4066376-0 gnd rswk-swf Zell-Adhäsionsmolekül (DE-588)4300852-5 gnd rswk-swf Down-Syndrom (DE-588)4012849-0 gnd rswk-swf Signaltransduktion (DE-588)4318717-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Wirbeltiere (DE-588)4066376-0 s Down-Syndrom (DE-588)4012849-0 s Zell-Adhäsionsmolekül (DE-588)4300852-5 s Signaltransduktion (DE-588)4318717-1 s DE-188 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028676617&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Sachse, Sonja Maria Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 Wirbeltiere (DE-588)4066376-0 gnd Zell-Adhäsionsmolekül (DE-588)4300852-5 gnd Down-Syndrom (DE-588)4012849-0 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| subject_GND | (DE-588)4066376-0 (DE-588)4300852-5 (DE-588)4012849-0 (DE-588)4318717-1 (DE-588)4113937-9 |
| title | Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 |
| title_auth | Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 |
| title_exact_search | Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 |
| title_full | Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 vorgelegt von: Sonja Maria Sachse aus Berlin |
| title_fullStr | Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 vorgelegt von: Sonja Maria Sachse aus Berlin |
| title_full_unstemmed | Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 vorgelegt von: Sonja Maria Sachse aus Berlin |
| title_short | Novel signaling functions of the neuronal cell adhesion molecules DSCAM amd DSCAML1 |
| title_sort | novel signaling functions of the neuronal cell adhesion molecules dscam amd dscaml1 |
| topic | Wirbeltiere (DE-588)4066376-0 gnd Zell-Adhäsionsmolekül (DE-588)4300852-5 gnd Down-Syndrom (DE-588)4012849-0 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| topic_facet | Wirbeltiere Zell-Adhäsionsmolekül Down-Syndrom Signaltransduktion Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028676617&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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