TSSK1β inhibits the Hippo pathway effector protein YAP to suppress cell proliferation
- 주제(키워드) Hippo pathway , LATS1/2 , YAP , TSSK1β , Cell proliferation , TEAD , Cancer
- 주제(DDC) 570
- 발행기관 아주대학교
- 지도교수 모정순
- 발행년도 2021
- 학위수여년월 2021. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000031171
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The Hippo signaling pathway, which is involved in the regulation of cell proliferation, and an important tumor suppressor. The central regulators of the Hippo signaling pathway include the transcriptional co-activators YAP /TAZ which are negatively regulated by LATS1/2 kinases. The phosphorylation of YAP/TAZ promote their cytoplasmic sequestration where they bind with 14-3-3 protein or undergo proteasomal degradation. Dysregulation of Hippo pathway promotes the oncogenic properties of YAP/TAZ and improves cancer progression. However, the potential of YAP as a therapeutic target for cancer has not been well studied. To identify kinases-induced the phosphorylation of YAP, we screened diverse kinases which belong to CAMK family. This study demonstrated that TSSK1β promoted YAP phosphorylation partially through the LATS1/2-dependent or –independent mechanisms. Furthermore, TSSK1β consequently inhibits cell proliferation and anchorage-independent growth in LATS1/2 DKO MEFs. In summary, our findings suggest that TSSK1β can suppress YAP/TAZ-mediated cancer cell proliferation. Thus, TSSK1β can be a potential therapeutic target carcinogenesis.
more목차
I. INTRODUCTION 1
II.MATERIALSAND METHODS 9
A. Reagents and antibodies 9
B. Cell culture and transfection 9
C. Retroviral infection and generation of stable cell lines 9
D. Western blotting and immunoprecipitation (IP) 10
E. Recombinant protein purification 10
F. Luciferase assay 11
G. In vitro kinase assay 11
H. Immunocytochemistry (ICC) 11
I. RNA isolation and quantitative real-time polymerase chain reaction (RT-qPCR) 12
J. Colony formation assay 13
K. Sulforhodamine B (SRB) assay 13
L. Clonogenic growth assay 13
M. Generation of knock-out cells 14
N. Statistical analysis 14
III. RESULTS 15
A. TSSK1β promotes YAP phosphorylation 15
B. TSSK1β drives YAP inactivation and cytoplasmic retention 18
C. TSSK1β induces YAP phosphorylation via LATS-dependent or -independent manners 21
D. TSSK1β inhibits cell proliferation and oncogenic potential of LATS 1/2 DKO MEFs 25
IV. DISCUSSION 28
V. CONCLUSION 31
VI. REFERENCE 32
