Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae
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2002
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| 100 | 1 | |a Finkbeiner, Martin G. |d 1970- |e Verfasser |0 (DE-588)124718825 |4 aut | |
| 245 | 1 | 0 | |a Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae |c von Martin G. Finkbeiner |
| 264 | 1 | |c 2002 | |
| 300 | |a [6], 110 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a München, Univ., Diss., 2003 | ||
| 650 | 4 | |a Saccharomyces cerevisiae - Meiose - Plasmamembran - Lipide - Bindeproteine | |
| 650 | 0 | 7 | |a Lipide |0 (DE-588)4035873-2 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Plasmamembran |0 (DE-588)4067550-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Saccharomyces cerevisiae |0 (DE-588)4178812-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Bindeproteine |0 (DE-588)4276933-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Meiose |0 (DE-588)4169347-4 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Saccharomyces cerevisiae |0 (DE-588)4178812-6 |D s |
| 689 | 0 | 1 | |a Meiose |0 (DE-588)4169347-4 |D s |
| 689 | 0 | 2 | |a Plasmamembran |0 (DE-588)4067550-6 |D s |
| 689 | 0 | 3 | |a Lipide |0 (DE-588)4035873-2 |D s |
| 689 | 0 | 4 | |a Bindeproteine |0 (DE-588)4276933-4 |D s |
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-010410903 | |
Datensatz im Suchindex
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| _version_ | 1823052598682320896 |
| adam_text |
TABLE OF CONTENTS
SUMMARY 1
ZUSAMMENFASSUNG 2
ABBREVIATIONS 4
1. INTRODUCTION 6
1.1 SEX AND THE SINGLE CELL: MEIOSIS IN YEAST 7
SPECIAL FEATURES OF THE MEIOTIC CELL CYCLE 8
MEIOSIS IN
SACCHAROMYCES CEREVISIAE 9
1.2 PROPHASE OF MEIOSIS I: RECOMBINATION EVENTS DURING ASSEMBLY 10
AND DISASSEMBLY OF THE SYNAPTONEMAL COMPLEX
THE RECOMBINATION PATHWAY IN MEIOSIS 11
CHROMOSOME MORPHOLOGY IN MEIOTIC PROPHASE 14
THE SYNAPTONEMAL COMPLEX 15
FUNCTION OF THE SYNAPTONEMAL COMPLEX 17
T .3 BUILDING NEW CELLS WITHIN OLD ONES: THE MORPHOGENETIC 18
PROGRAM AT THE END OF MEIOSIS INVOLVES
DE NOVO
MEMBRANE FORMATION
SPINDLE POLE BODIES MEET AN ADDITIONAL TASK IN MEIOSIS 18
DISCOVERY OF PROTEINS LOCALIZING SPECIFICALLY TO 20
A SUBSTRUCTURE OF THE PROSPORE MEMBRANE
DURING MEIOSIS MEMBRANE VESICLES OF THE SECRETORY 23
PATHWAY ARE REDIRECTED TO THE SPINDLE POLE BODY
AIMS OF THIS STUDY 24
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/967768551
2. RESULTS 25
2.1 STUDIES ON THE SYNAPTONEMAL COMPLEX PROTEIN ZIP1 P 26
EXPRESSION OF ZIP1 P IN A MEIOTIC TIME COURSE 26
ZIP1 P LOCALIZATION ALONG SYNAPSED CHROMOSOMES AND 28
TO POLYCOMPLEXES IN MEIOTIC SPREAD EXPERIMENTS
TWO HYBRID INTERACTIONS AMONG ZIP1 P DOMAINS 29
SEARCH FOR A ZIP1 P INTERACTING PARTNER WITH BIOCHEMICAL 31
AND GENETICAL METHODS
2.2 BIOCHEMICAL ANALYSIS OF THE MEIOSIS SPECIFIC PROTEIN SSP1 P 33
EXPRESSION OF SSPL P IN A MEIOTIC TIME COURSE 33
FRACTIONATION OF SSPLP REVEALS MODIFICATION OF THE PROTEIN 34
DEPHOSPHORYLATION EXPERIMENTS PROVE THAT SSP1 P 36
IS A PHOSPHOPROTEIN
GEL FILTRATION EXPERIMENTS OF SSP1 P 37
2.3 IDENTIFICATION OF SSP1 P AS ANOTHER COMPONENT OF THE 38
LEADING EDGE PROTEIN (LEP) COAT
SSP1 P LOCALIZES ALL ALONG THE PROSPORE MEMBRANE 38
SSP1 P FORMS RING-LIKE STRUCTURES IN MEIOTIC SPREAD 40
EXPERIMENTS
COLOCALIZATION OF SSP1 P WITH LEP COAT COMPONENTS 41
LOCALIZATION OF SSP1 P TO THE SPINDLE POLE BODY AND TO 41
THE LEP COAT DOES NOT DEPEND ON ADY3P
SSP1 P LOCALIZES TO THE LEP COAT AND ADDITIONALLY ALL 43
ALONG THE PROSPORE MEMBRANE
2.4 PHENOTYPIC ANALYSIS OF THE ASSPF DELETION MUTANT 44
PROSPORE MEMBRANE FORMATION AND SPORE WALL 45
MATURATION IN THE WILD TYPE STRAIN
IN THE
SSPL MUTANT PROSPORE MEMBRANE SHAPING 47
IS ABNORMAL
ATYPICAL BODIES SURROUNDED BY TWO MEMBRANE SYSTEMS 47
ACCUMULATE IN THE SSP1 MUTANT
THE
SSP1 MUTANT LOSES VIABILITY IN THE COURSE 50
OF SPORULATION
DELETION OF
SSP1 DOES NOT BLOCK TRANSCRIPTION OF LATE 50
MEIOTIC GENES
2.5 LIPID BINDING PROPERTIES AND MEMBRANE LOCALIZATION OF SSP1 P 53
FULL LENGTH SSP1 P BINDS TO PHOSPHATIDYLSERINE, CARDIO- 54
LIPIN AND TO ALL PHOSPHATIDYLINOSITOLPHOSPHATES (PIPS)
PURIFICATION OF SSP1 P SUBDOMAINS 56
THE N-TERMINUS OF SSP1 P COMPRISES THE PIP BINDING 58
ACTIVITY
MITOTICALLY OVEREXPRESSED SSP1 P-GFP LOCALIZES TO SITES 61
OF SECRETORY VESICLE FUSION AT THE BUD PLASMA MEMBRANE
SSP1 P HAS CYTOTOXIC EFFECTS WHEN IT IS MITOTICALLY 63
OVEREXPRESSED IN YEAST
MAPPING OF THE REGION IN SSPLP RESPONSIBLE FOR 65
ADY3P BINDING
2.6 ANALYSIS OF MUTANT PHENOTYPES FOR A
SPO70 AND ASPO71 STRAINS 68
EPISTASIS EXPERIMENTS COMBINING MUTANTS OF
SSP1, 69
SPO70 AND SPO71
IMMATURE SPORES CONTAINING CYTOPLASM ARE STILL FORMED 71
IN THE ASPO70 AND ASPO71 DELETION MUTANTS
3. MATERIALS AND METHODS 72
3.1 GENERAL CONSIDERATIONS 73
3.2 GENE TAGGING AND DISRUPTION WITH A PCR BASED CASSETTE 73
MECHANISM
3.3 SPORULATION CONDITIONS
74
3.4 TWO HYBRID ASSAY
74
3.5 TWO HYBRID SCREEN
75
3.6 YEAST PLASMID RESCUE 76
3.7 PROTEIN OVEREXPRESSION IN YEAST 76
3.8 PREPARING TCA EXTRACTS FROM YEAST 77
3.9 GLASS BEAD LYSIS 78
3.10 FRACTIONATION OF YEAST EXTRACTS 78
3.11 PROTEASE INHIBITORS 78
3.12 PURIFICATION OF PROTEIN COMPLEXES USING THE PROA TAG 79
3.13 DEPHOSPHORYLATION ASSAY 80
3.14 GEL FILTRATION 80
3.15 PROTEIN-LIPID OVERLAY ASSAY 81
3.16 PREPARATION AND PURIFICATION OF POLYCLONAL RABBIT ANTIBODIES 81
3.17 IMMUNOFLUORESCENCE MICROSCOPY 82
3.18 MEIOTIC SPREADS 83
3.19 ELECTRON MICROSCOPY 84
3.20 OVEREXPRESSION OF YEAST PROTEINS IN BACTERIA AND PURIFICATION 85
3.21 NORTHERN BLOT EXPERIMENTS 86
3.22 LIST OF ANTIBODIES 87
3.23 TABLE OF PLASMIDS 88
3.24 TABLE OF YEAST STRAINS 90
4. DISCUSSION 91
4.1 SSPLP IS A NOVEL LEADING EDGE PROTEIN COAT COMPONENT 92
4.2 DELETION OF
SSP1 RESULTS IN MISSHAPING OF THE PROSPORE MEMBRANE 94
4.3 LIPID BINDING OF SSPLP ANCHORS THE LEP COAT IN THE MEMBRANE 97
4.4 SSP1 P MIGHT BE INVOLVED IN MEMBRANE VESICLE TRAFFICKING 99
ACKNOWLEDGEMENTS 103
LEBENSLAUF 104
BIBLIOGRAPHY 105 |
| any_adam_object | 1 |
| author | Finkbeiner, Martin G. 1970- |
| author_GND | (DE-588)124718825 |
| author_facet | Finkbeiner, Martin G. 1970- |
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| indexdate | 2025-02-03T16:57:40Z |
| institution | BVB |
| language | English |
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| physical | [6], 110 Bl. Ill., graph. Darst. |
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| spellingShingle | Finkbeiner, Martin G. 1970- Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae Saccharomyces cerevisiae - Meiose - Plasmamembran - Lipide - Bindeproteine Lipide (DE-588)4035873-2 gnd Plasmamembran (DE-588)4067550-6 gnd Saccharomyces cerevisiae (DE-588)4178812-6 gnd Bindeproteine (DE-588)4276933-4 gnd Meiose (DE-588)4169347-4 gnd |
| subject_GND | (DE-588)4035873-2 (DE-588)4067550-6 (DE-588)4178812-6 (DE-588)4276933-4 (DE-588)4169347-4 (DE-588)4113937-9 |
| title | Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae |
| title_auth | Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae |
| title_exact_search | Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae |
| title_full | Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae von Martin G. Finkbeiner |
| title_fullStr | Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae von Martin G. Finkbeiner |
| title_full_unstemmed | Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae von Martin G. Finkbeiner |
| title_short | Ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in S. cerevisiae |
| title_sort | ssp1p is a lipid binding protein involved in shaping of the prospore membrane during meiosis in s cerevisiae |
| topic | Saccharomyces cerevisiae - Meiose - Plasmamembran - Lipide - Bindeproteine Lipide (DE-588)4035873-2 gnd Plasmamembran (DE-588)4067550-6 gnd Saccharomyces cerevisiae (DE-588)4178812-6 gnd Bindeproteine (DE-588)4276933-4 gnd Meiose (DE-588)4169347-4 gnd |
| topic_facet | Saccharomyces cerevisiae - Meiose - Plasmamembran - Lipide - Bindeproteine Lipide Plasmamembran Saccharomyces cerevisiae Bindeproteine Meiose Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010410903&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT finkbeinermarting ssp1pisalipidbindingproteininvolvedinshapingoftheprosporemembraneduringmeiosisinscerevisiae |