Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum:
Gespeichert in:
| Beteilige Person: | |
|---|---|
| Format: | Hochschulschrift/Dissertation Microform Buch |
| Sprache: | Englisch |
| Veröffentlicht: |
Köln
2018
|
| Schlagwörter: | |
| Links: | http://d-nb.info/1163746835/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030667822&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| Umfang: | 2 Mikrofiches (VI, 115 Seiten) Illustrationen |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV045280235 | ||
| 003 | DE-604 | ||
| 005 | 00000000000000.0 | ||
| 007 | hu|uuuuuuuucu | ||
| 008 | 181109s2018 gw a||| am||| 00||| eng d | ||
| 015 | |a 18,H09 |2 dnb | ||
| 016 | 7 | |a 1163746835 |2 DE-101 | |
| 035 | |a (OCoLC)1136260263 | ||
| 035 | |a (DE-599)DNB1163746835 | ||
| 040 | |a DE-604 |b ger |e rda | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c XA-DE | ||
| 049 | |a DE-355 | ||
| 100 | 1 | |a Fischer, Sarah |d 1990- |e Verfasser |0 (DE-588)1162524367 |4 aut | |
| 245 | 1 | 0 | |a Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum |c vorgelegt von Sarah Fischer |
| 264 | 1 | |a Köln |c 2018 | |
| 300 | |a 2 Mikrofiches (VI, 115 Seiten) |b Illustrationen | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b h |2 rdamedia | ||
| 338 | |b he |2 rdacarrier | ||
| 340 | |f 24x | ||
| 502 | |b Dissertation |c Universität zu Köln |d 2018 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u http://d-nb.info/1163746835/04 |3 Inhaltsverzeichnis |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030667822&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-030667822 | |
Datensatz im Suchindex
| _version_ | 1819277586121883648 |
|---|---|
| adam_text | TABLE OF CONTENTS
TABLE OF
CONTENTS......................................................................................................
IV
1
INTRODUCTION...........................................................................................................
1
1.1 THE MODEL ORGANISM DICTYOSTELIUM
DISCOIDEUM......................................................1
1.2 PROTEIN TURNOVER
MECHANISMS...................................................................................3
1.2.1 THE UBIQUITIN-PROTEASOME SYSTEM
(UPS)........................................................3
1.2.2
AUTOPHAGY........................................................................................................
5
1.2.3 CROSSTALK BETWEEN AUTOPHAGY AND THE
UPS..................................................14
1.3 KEY AIMS OF THIS
WORK..............................................................................................
16
1.4
PUBLICATIONS.............................................................................................................16
2
RESULTS..................................................................................................................17
2.1 GENERATION AND VERIFICATION OF ATG12* AND ATG12716 KNOCK-OUT
STRAINS
........
17
2.2 GENERATION OF ATG 12-SPECIFIC MONOCLONAL
ANTIBODIES.........................................19
2.2.1 PURIFICATION OF RECOMBINANT GST-TAGGED
ATG12.........................................19
2.2.2 THE IMMUNISATION OF MICE WITH RECOMBINANT ATG12 RESULTED IN
SPECIFIC
ANTIBODIES.......................................................................................................20
2.3 MUTANT STRAINS SHOW SEVERE DEVELOPMENTAL
DEFECTS.............................................23
2.4 CELL AND SPORE VIABILITY ARE DECREASED IN ATG 12 AND ATG 16
DEFICIENT CELLS
.....
25
2.5 GROWTH BEHAVIOUR IS ALTERED IN ATG12 AND ATG16 KNOCK-OUT STRAINS
................
26
2.5.1 CELL GROWTH IN LIQUID CULTURE IS SLOWED DOWN IN STRAINS LACKING
ATG12 AND
ATG
16............................................................................................................
26
2.5.2 GROWTH ON K. AEROGENES IS OPPOSITELY AFFECTED IN ATG12* AND ATG16*
CELLS................................................................................................................27
2.6 AUTOLYSOSOME FORMATION IS DISTURBED IN MUTANT STRAINS
......................................
28
2.7 PHAGOCYTOSIS OF TRITC-LABELLED YEAST IS LESS EFFICIENT IN MUTANT
STRAINS
...........
30
2.8 ATG12 AND ATG16 DEFICIENCY RESULTS IN IMPAIRED MACROPINOCYTOSIS
................
31
2.9 PROTEIN HOMEOSTASIS IS DISTURBED IN KNOCK-OUT
STRAINS.........................................32
2.9.1 GLOBAL PROTEIN UBIQUITINATION AND PROTEIN AGGREGATES ARE INCREASED
IN
ATG 12 AND ATG 16 DEFICIENT
CELLS............................................................... 32
2.9.2 PROTEASOMAL ACTIVITY IS SIGNIFICANTLY DECREASED IN MUTANT STRAINS
..............
34
2.10
RN ASEQ
ANALYSIS OF AX2, ATG 12*, ATG 16* AND ATG 12716* CELLS
.....................
35
2.10.1 MASSIVE TRANSCRIPTIONAL CHANGES OCCUR IN MUTANT STRAINS
..........................
35
2.10.2 ATG12 AND ATG16 DEFICIENCY RESULTS IN UP-REGULATION OF OTHER CORE
AUTOPHAGY
GENES...........................................................................................
41
2.10.3 GENE ONTOLOGY ANALYSIS REVEALS DIFFERENTIALLY REGULATED ENRICHED
GENE SETS
IN ATG12 ATG16 AND ATG12716* STRAINS
..............................................
43
2.11 SUMMARY OF ANALYSED CELLULAR PROCESSES IN KNOCK-OUT
STRAINS.............................45
3
DISCUSSION.........................................................................................................
46
3.1 IN WILD-TYPE CELLS ONLY ATG12-ATG5 CONJUGATES, BUT NO ATG12 MONOMERS
COULD
BE
DETECTED.............................................................................................................
46
3.2 ATG12 IS PRESENT IN VESICULAR STRUCTURES THROUGHOUT THE CYTOPLASM
...................
47
3.3 ATG12 AND/OR ATG16 DEFICIENCY CAUSED DISTINCT
PHENOTYPES............................49
3.4 ATG12 AND ATG16 ARE REQUIRED FOR EFFICIENT AUTOPHAGOSOME FORMATION
AND
MATURATION...............................................................................................................
51
3.5 PROTEIN HOMEOSTASIS IS IMPAIRED IN ATG12 AND ATG16 DEFICIENT STRAINS
...........
52
3.6 MASSIVE TRANSCRIPTIONAL CHANGES OCCURRED IN ATG12 AND/OR ATG16
DEFICIENT
STRAINS......................................................................................................................54
4 MATERIALS AND
METHODS.....................................................................................56
4.1
MATERIALS..................................................................................................................56
4.1.1
STRAINS............................................................................................................
56
4.1.2
ANTIBODIES......................................................................................................57
4.1.3
PLASMIDS........................................................................................................
57
4.1.4
ANTIBIOTICS.......................................................................................................57
4.1.5
OLIGONUCLEOTIDES............................................................................................58
4.1.6
CHEMICALS......................................................................................................58
4.1.7
ENZYMES........................................................................................................
59
4.1.8
KITS.................................................................................................................
60
4.1.9 MEDIA, BUFFERS AND AGAR
PLATES......................................................................60
4.2
METHODS..................................................................................................................
62
4.2.1 MICROBIOLOGICAL
METHODS................................................................................
62
4.2.2 CELL BIOLOGICAL
METHODS.................................................................................64
4.2.3 MOLECULAR BIOLOGICAL
METHODS.......................................................................
68
4.2.4 BIOCHEMICAL AND IMMUNOLOGICAL METHODS
...................................................
78
4.2.5 IMMUNOFLUORESCENCE
STUDIES........................................................................
85
4.2.6 STATISTICS AND REPRODUCIBILITY
.........................................................................
87
5
REFERENCES..........................................................................................................88
6
ABBREVIATIONS....................................................................................................104
7 LIST OF
FIGURES....................................................................................................107
8 LIST OF
TABLES......................................................................................................109
9
SUMMARY...........................................................................................................110
10
ZUSAMMENFASSUNG...........................................................................................I
L L
11
APPENDIX...........................................................................................................112
12
ERKLAERUNG...........................................................................................................114
13 CURRICULUM
VITAE..............................................................................................115
|
| any_adam_object | 1 |
| author | Fischer, Sarah 1990- |
| author_GND | (DE-588)1162524367 |
| author_facet | Fischer, Sarah 1990- |
| author_role | aut |
| author_sort | Fischer, Sarah 1990- |
| author_variant | s f sf |
| building | Verbundindex |
| bvnumber | BV045280235 |
| ctrlnum | (OCoLC)1136260263 (DE-599)DNB1163746835 |
| format | Thesis Microfilm Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01334nam a2200337 c 4500</leader><controlfield tag="001">BV045280235</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">00000000000000.0</controlfield><controlfield tag="007">hu|uuuuuuuucu</controlfield><controlfield tag="008">181109s2018 gw a||| am||| 00||| eng d</controlfield><datafield tag="015" ind1=" " ind2=" "><subfield code="a">18,H09</subfield><subfield code="2">dnb</subfield></datafield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">1163746835</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)1136260263</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)DNB1163746835</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rda</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="044" ind1=" " ind2=" "><subfield code="a">gw</subfield><subfield code="c">XA-DE</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-355</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Fischer, Sarah</subfield><subfield code="d">1990-</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)1162524367</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum</subfield><subfield code="c">vorgelegt von Sarah Fischer</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Köln</subfield><subfield code="c">2018</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">2 Mikrofiches (VI, 115 Seiten)</subfield><subfield code="b">Illustrationen</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">h</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">he</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="340" ind1=" " ind2=" "><subfield code="f">24x</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="b">Dissertation</subfield><subfield code="c">Universität zu Köln</subfield><subfield code="d">2018</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">B:DE-101</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://d-nb.info/1163746835/04</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030667822&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="943" ind1="1" ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-030667822</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV045280235 |
| illustrated | Illustrated |
| indexdate | 2024-12-20T18:22:56Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-030667822 |
| oclc_num | 1136260263 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | 2 Mikrofiches (VI, 115 Seiten) Illustrationen |
| publishDate | 2018 |
| publishDateSearch | 2018 |
| publishDateSort | 2018 |
| record_format | marc |
| spellingShingle | Fischer, Sarah 1990- Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum |
| subject_GND | (DE-588)4113937-9 |
| title | Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum |
| title_auth | Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum |
| title_exact_search | Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum |
| title_full | Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum vorgelegt von Sarah Fischer |
| title_fullStr | Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum vorgelegt von Sarah Fischer |
| title_full_unstemmed | Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum vorgelegt von Sarah Fischer |
| title_short | Functional analysis of the core autophagy protein ATG12 in Dictyostelium discoideum |
| title_sort | functional analysis of the core autophagy protein atg12 in dictyostelium discoideum |
| topic_facet | Hochschulschrift |
| url | http://d-nb.info/1163746835/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030667822&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT fischersarah functionalanalysisofthecoreautophagyproteinatg12indictyosteliumdiscoideum |