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A study on the essential role of casein kinase gamma 1 and 3 in necroptosis : 세포괴사 조절 인자로서 카제인 키나아제 1 감마에 대한 연구
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 정용근 | - |
dc.contributor.author | 이송이 | - |
dc.date.accessioned | 2018-11-12T00:54:09Z | - |
dc.date.available | 2018-11-12T00:54:09Z | - |
dc.date.issued | 2018-08 | - |
dc.identifier.other | 000000153099 | - |
dc.identifier.uri | https://hdl.handle.net/10371/143013 | - |
dc.description | 학위논문 (박사)-- 서울대학교 대학원 : 자연과학대학 생명과학부, 2018. 8. 정용근. | - |
dc.description.abstract | Upon necroptosis activation, receptor interacting serine/threonine kinase
(RIPK) 1 and RIPK3 form a necrosome complex with pseudokinase mixed lineage kinase-like (MLKL). Although protein phosphorylation is a key event for the activation of RIPK1 and RIPK3 in response to necroptosis signal, relatively little is known about other factors that can regulate the activity of those kinases or necrosome formation. In a gain-of-function screen with 650 kinases and 120 phosphatases, I identified that casein kinase 1 gamma (CK1γ) as a crucial regulator of necroptosis. Here I show that the downregulation of CK1γ1 and CK1γ3, either by a chemical inhibitor or knockdown in cells, reduced TNFα-induced necroptosis. Conversely, ectopic expression of CK1γ1 or CK1γ3 exacerbated necroptosis, but not apoptosis. Like RIPK1 and RIPK3, CK1γ1 was cleaved at Asp434 by caspase-8 during apoptosis, while it was increased in response to necroptosis. CK1γ1 and CK1γ3 formed a protein complex with each other and were recruited into the necrosome harboring RIPK1, RIPK3 and MLKL. Especially, autophosphorylated form of CK1γ3 at Ser344/345 was detected in the necrosome and was required to mediate the necroptosis. In addition, CK1γ phosphorylates RIPK3 in vitro and a CK1γ-specific inhibitor Gi reduced the phosphorylation of MLKL at Ser358, as well as the formation of MLKL oligomers, and rescued mice from TNFα-induced systemic inflammatory response syndrome (SIRS). Collectively, these data suggest that CK1γ1 and CK1γ3 are required for promoting cell death progression by regulating the formation of necrosome through RIPK3. | - |
dc.description.tableofcontents | 1. INTRODUCTION . 1
2. MATERIAL AND METHOD 5 2.1. Collection of cDNAs and GOF screening 5 2.2. Cell culture and stable cell lines . 5 2.3. Reagents and antibodies . 6 2.4. Plasmid construction . 7 2.5. shRNA-mediated knockdown . 8 2.6. Cell death assays 9 2.7 Immunoprecipitation and subcellular fractionation . 10 2.8. In vitro binding assay 11 2.9. In vitro kinase assay 11 2.10. Statistical analysis 12 3. RESULTS 13 3.1. CK1γ identified in a functional screen is involved in mediating TSI-induced necroptosis . 13 3.2. CK1γ1 is cleaved by caspase-8 in vitro and in apoptotic cell . 15 3.3. CK1γ1 and CK1γ3 are accumulated in response to TSI during necroptosis 17 3.4. CK1γ is a component of necrosome harboring active RIPK1, RIPK3 and MLKL. 19 3.5. CK1γ is autophosphorylated and phosphorylates RIPK3 to activate it in vitro . 21 3.6. CK1γ3 is autophosphorylated at Ser344 and 345 to exert its activity in necroptosis . 23 3.7. Gi protects mice in a TNFα-induced systemic inflammatory response syndrome (SIRS) model 25 4. DISCUSSION 64 5. REFERENCES 70 6. 국문초록 80 | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject.ddc | 570 | - |
dc.title | A study on the essential role of casein kinase gamma 1 and 3 in necroptosis | - |
dc.title.alternative | 세포괴사 조절 인자로서 카제인 키나아제 1 감마에 대한 연구 | - |
dc.type | Thesis | - |
dc.contributor.AlternativeAuthor | Lee, Song-Yi | - |
dc.description.degree | Doctor | - |
dc.contributor.affiliation | 자연과학대학 생명과학부 | - |
dc.date.awarded | 2018-08 | - |
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