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ARD1-mediated Aurora kinase A acetylation promotes cancer cell proliferation and migration

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dc.contributor.advisorKyu-Won Kim-
dc.contributor.author보탐투이루-
dc.date.accessioned2017-10-27T17:01:03Z-
dc.date.available2017-10-27T17:01:03Z-
dc.date.issued2017-08-
dc.identifier.other000000145721-
dc.identifier.urihttps://hdl.handle.net/10371/137017-
dc.description학위논문 (박사)-- 서울대학교 대학원 약학대학 약학과, 2017. 8. Kyu-Won Kim.-
dc.description.abstractAurora kinase A (AuA) is a prerequisite for centrosome maturation, separation, and mitotic spindle assembly, thus, it is essential for cell cycle regulation. Overexpression of AuA is implicated in poor prognosis of many types of cancer. However, the regulatory mechanisms underlying the functions of AuA are still not fully understood. Here, we report that AuA colocalizes with arrest defective protein 1 (ARD1) acetyltransferase during cell division and cell migration. Additionally, AuA is acetylated by ARD1 at lysine residues at positions 75 and 125. The double mutations at K75/K125 abolished the kinase activity of AuA. Moreover, the double mutant AuA exhibited diminished ability to promote cell proliferation and cell migration. Mechanistic studies revealed that AuA acetylation at K75/K125 promoted cell proliferation via activation of cyclin E/CDK2 and cyclin B1. In addition, AuA acetylation stimulated cell migration by activating the p38/AKT/MMP-2 pathway. These findings indicate that ARD1-mediated acetylation of AuA enhances cell proliferation and migration, and probably contributes to cancer development.-
dc.description.tableofcontentsINTRODUCTION 1
I. Protein acetylation 1
II. ARD1 5
III. Aurora kinases 9
1. Aurora kinase family 9
2. Structure of Aurora kinases 10
3. Subcellular localizations and functions of Aurora kinases 12
4. Activation and degradation of Aurora kinase A 15
5. Mitoticfunction of Aurora kinase A 18
6. Non-mitotic function of Aurora kinase A 22

PURPOSE OF THIS STUDY 25
MATERIALS AND METHODS 27

1. Antibodies 27
2. Plasmid construction .28
3. Cell culture 29
4. Cell synchronization 29
5. Immunoflourescence 30
6. Immunoblotting and immunoprecipitation 31
7. In vitro acetylation assay 32
8. ATP-binding assay 33
9. ATPase activity assay 33
10. Protein sequence alignment 34
11. Cell proliferation assay 34
12. Clonogenic assay 35
13. Cell cycle analysis 35
14. Wound healing assay 36
15. Transwell invasion assay 36
16. Statistical analysis .37

RESULTS 38
1. ARD1 has centrosome-like localization during cell cycle progression 38
2. ARD1 colocalizes with Aurora kinase A during cell cycle progression 41
3. ARD1 colocalizes with Aurora kinase A during cell migration 43
4. Aurora kinase A is acetylated by ARD1 45
5 Acetylation of Aurora kinase A is dependent on ARD1 catalytic activity 49
6 Aurora kinase A is acetylated at at lysine residues at position at 75 and 125 52
7 Double mutation at lysine 75 and 125 abolished acetylation of AuA 56
8 Acetylation of Aurora kinase A increases the phosphorylation of Aurora kinase A 60
9 Acetylation of Aurora kinase A increases the kinase activity of Aurora kinase A 62
10 Acetylation of Aurora kinase A promotes cell proliferation 68
11 Acetylation of Aurora kinase A increases G2/M cells population 73
12 Acetylation of Aurora kinase A promotes cell proliferation by increasing expression level of cell cycle check point proteins 76
13 Acetylation of Aurora kinase A increases during cell migration 79
14 Acetylation of Aurora kinase A promotes cell migration 81
15 Acetylation of Aurora kinase A promotes cell migration via p38/AKT/MMP2 pathway 84
DISCUSSION 87
REFERENCES 94
ABSTRACT IN KOREAN (국문초록) 114
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dc.formatapplication/pdf-
dc.format.extent2257959 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectARD1-
dc.subjectAurora kinase A-
dc.subjectlysine acetylation-
dc.subjectcell proliferation-
dc.subjectcell migration-
dc.subject.ddc615-
dc.titleARD1-mediated Aurora kinase A acetylation promotes cancer cell proliferation and migration-
dc.typeThesis-
dc.description.degreeDoctor-
dc.contributor.affiliation약학대학 약학과-
dc.date.awarded2017-08-
Appears in Collections:
College of Pharmacy (약학대학)Dept. of Pharmacy (약학과)Theses (Ph.D. / Sc.D._약학과)
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