Characterization of the budding yeast microtubule polymerase Stu2:
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| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XII, 90 S. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Characterization of the budding yeast microtubule polymerase Stu2 |c Marija Podolski |
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Datensatz im Suchindex
| _version_ | 1804152044598067200 |
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| adam_text | CONTENTS
LIST OF FIGURES IX
LIST OF TABLES XI
GLOSSARY XIII
1 INTRODUCTION 1
1.1 HISTORY OF CELL DIVISION AND MICROTUBULE RESEARCH 1
1.2 TUBULIN AND MICROTUBULE STRUCTURE 2
1.3 DYNAMIC INSTABILITY OF MICROTUBULES 4
1.4 TUBULIN COMPLEXITY AND POST-TRANSLATIONAL MODIFICATIONS 7
1.5 REGULATION OF MICROTUBULE DYNAMICS IN CELLS 8
1.6 XMAP215/DISL FAMILY OF MICROTUBULE POLYMERASES 9
1.7 STU2, THE BUDDING YEAST MEMBER OF XMAP215/ DISL FAMILY ... 12
1.8 YEAST AS A MODEL SYSTEM FOR STUDYING MICROTUBULE DYNAMICS ... 13
2 AIM OF THIS WORK 15
3 EFFECTS OF STU2 ON PORCINE BRAIN MICROTUBULES 17
3.1 STU2 BINDS TO THE MICROTUBULE ENDS 17
3.2 BINDING OF STU2 TO THE MICROTUBULE IS SALT DEPENDENT 19
3.3 STU2 DEPOLYMERIZES GMPCPP STABILIZED MICROTUBULES IN ABSENCE
OF TUBULIN 20
3.4 STU2 LOCALIZES TO A SPECIFIC REGION OF THE GROWING MICROTUBULE ...
20
3.5 STU2 IS A WEAK POLYMERASE OF PORCINE MICROTUBULES 23
3.6 DISCUSSION 23
V
HTTP://D-NB.INFO/1049406400
CONTENTS
4 DEVELOPMENT OF THE TOG COLUMN 29
4.1 MAKING THE TOG COLUMN 30
4.2 TOG COLUMN BINDS PORCINE BRAIN TUBULIN 31
4.3 FUNCTIONAL TUBULIN CAN BE PURIFIED FROM BUDDING YEAST 31
4.4 CYCLING OF YEAST TUBULIN IN HIGH MOLARITY SALT BUFFER 35
4.5 YEAST TUBULIN IS FUNCTIONAL AND FORMS MICROTUBULES AT LOW CONCEN
TRATIONS 37
4.6 DISCUSSION 38
5 EFFECTS OF STU2 ON BUDDING YEAST MICROTUBULES 43
5.1 COMPARISON OF STU2 S EFFECT ON YEAST AND PIG MICROTUBULES .... 43
5.2 COMPARISON OF STU2 AND XMAP215 47
5.3 THE UNSTEADINESS OF GROWTH IN PRESENCE OF XMAP215 49
5.4 DISCUSSION 52
6 CONCLUSIONS 59
7 OUTLOOK 61
7.1 RESCUE PROMOTION BY STU2 61
7.2 TECHNICAL IMPROVEMENTS 61
7.3 POLYMERASE MUTANTS 62
7.4 YEAST TUBULIN MUTANTS AND THE EFFECTS OF POST-TRANSLATIONAL MODI
FICATIONS ON MICROTUBULE DYNAMICS 63
7.5 COMPARISON OF KIP3 AND KIFL8 ON THIER CONSPECIFIC TUBULIN .... 63
8 MATERIALS &: METHODS 65
8.1 PROTEIN EXPRESSION, PURIFICATION AND HANDLING 65
8.1.1 PURIFICATION OF STU2 65
8.1.2 SDS-PAGE 66
8.1.3 PROTEIN CONCENTRATION MEASUREMENTS 66
8.1.4 PORCINE BRAIN TUBULIN AND MICROTUBULE PREPARATION .... 66
8.1.5 CYCLING OF YEAST TUBULIN IN HIGH MOLARITY BUFFER 66
8.1.6 PURIFICATION OF TOGL AND TOG1/2-GST 67
8.1.7 COUPLING OF THE TOG DOMAIN TO THE NHS RESIN 68
VI
CONTENTS
8.1.8 MASS SPECTROMETRY OF YEAST TUBULIN CONTAMINANTS 68
8.2 EXPERIMENTAL SETUP 69
8.2.1 IMAGING AND ASSAY CONDITIONS 69
9 APPENDIX 71
REFERENCES 77
VII
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| bvnumber | BV041748273 |
| classification_rvk | WD 5050 |
| ctrlnum | (OCoLC)915431613 (DE-599)BSZ400349973 |
| discipline | Biologie |
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| indexdate | 2024-07-10T01:04:27Z |
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| physical | XII, 90 S. Ill., graph. Darst. |
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| spelling | Podolski, Marija 1984- Verfasser (DE-588)1047031175 aut Characterization of the budding yeast microtubule polymerase Stu2 Marija Podolski 2013 XII, 90 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Dresden, Techn. 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=027194720&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Podolski, Marija 1984- Characterization of the budding yeast microtubule polymerase Stu2 |
| subject_GND | (DE-588)4113937-9 |
| title | Characterization of the budding yeast microtubule polymerase Stu2 |
| title_auth | Characterization of the budding yeast microtubule polymerase Stu2 |
| title_exact_search | Characterization of the budding yeast microtubule polymerase Stu2 |
| title_full | Characterization of the budding yeast microtubule polymerase Stu2 Marija Podolski |
| title_fullStr | Characterization of the budding yeast microtubule polymerase Stu2 Marija Podolski |
| title_full_unstemmed | Characterization of the budding yeast microtubule polymerase Stu2 Marija Podolski |
| title_short | Characterization of the budding yeast microtubule polymerase Stu2 |
| title_sort | characterization of the budding yeast microtubule polymerase stu2 |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027194720&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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