Role of DNA binding in the regulation and function of ribosomal S6 kinase 2

DNA 结合在核糖体 S6 激酶 2 的调节和功能中的作用

• 批准号: BB/L010410/1
• 负责人: Ivan Gout
• 金额: $ 60.44万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL010410%2F1
• 关键词: Role; DNA; binding; regulation; function

Ribosomal protein S6 kinase (S6K) is a member of the AGC family of Ser Thr kinases which also includes PKA, PKB (Akt), PKCs etc. Biochemical and genetic studies in cell-based and animal models have provided evidence that S6K is a principal player in the regulation of cell growth, size and energy metabolism. Two major signal transduction pathways, phosphatidylinositide 3-kinases (PI3K) and mammalian target of rapamycin (mTOR), coordinate the activity of S6Ks in response to extracellular and intracellular stimuli, such as growth factors, mitogens, metabolites and nutrients. In an activate state, S6Ks translocate to discrete cellular compartments/multienzyme complexes, where they interact with and phosphorylate diverse substrates implicated in the regulation of translation, RNA processing, cytoskeletal rearrangement, cell growth and survival. A growing body of evidence links S6K signalling to various human pathologies, including diabetes, ageing and cancer. In mammalian cells, there are two isoforms of S6K, termed S6K1 and S6K2. Nucleocytoplasmic shuttling has been reported for both isoforms of S6Ks, but their nuclear functions are not well understood. This grant application is based on our novel findings which imply the role of S6K2 in the regulation of transcription. We provide the evidence that S6K2 possesses an AT-hook DNA-binding domain at the C-terminal autoinhibitory domain, which mediates specific interaction with DNA. We also found that the activity of S6K2 is induced by direct interaction with DNA in a dose-dependent manner. Furthermore, mutational studies of the AT-hook motif revealed its importance in the regulation of S6K2 pro-survival signalling. To further advance these original findings, we propose to study molecular mechanisms of S6K2-DNA interaction and an alternative model of S6K2 activation upon DNA binding by employing a range of biophysical, biochemical and cellular approaches. Our efforts will be also focused on exploring a cooperative mode of interaction between S6K2 and transcription factors in targeting promoter and enhancer regions of a specific set of genes, implicated in the regulation of S6K2-mediated cellular processes. Mammalian cell models and xenograft studies in nude mice will be employed to study the role of the S6K2 AT-hook motif in the regulation of transcription, ribosome biogenesis, cell survival and growth. It is expected that the proposed study will give deeper insight into nuclear functions of S6K2 and its implication in the regulation of transcription. Unique features of the S6K2-DNA interaction are not only of academic interest, but may also provide novel insights applicable to drug development.

核糖体蛋白S6激酶（ribosomalproteinS 6 kinase，S6 K）是丝氨酸-苏氨酸激酶（Ser-Thr kinases，AGC）家族的一员，该家族还包括PKA、PKB（Akt）、PKCs等。两种主要的信号传导途径，即磷脂酰肌醇3-激酶（PI 3 K）和哺乳动物雷帕霉素靶蛋白（mTOR），协调S6 KS对细胞外和细胞内刺激（例如生长因子、有丝分裂原、代谢物和营养素）的反应活性。在激活状态下，S6 K易位到离散的细胞区室/多酶复合物，在那里它们与涉及翻译、RNA加工、细胞骨架重排、细胞生长和存活的调节的不同底物相互作用并磷酸化。越来越多的证据将S6 K信号与各种人类病理联系起来，包括糖尿病，衰老和癌症。在哺乳动物细胞中，有两种S6 K亚型，称为S6 K1和S6 K2。核质穿梭已被报道为两种亚型的S6 Ks，但他们的核功能还没有得到很好的理解。这项拨款申请是基于我们的新发现，这意味着S6 K2在转录调控中的作用。我们提供的证据表明，S6 K2具有AT钩DNA结合域的C-末端的自抑制结构域，介导的特异性相互作用与DNA。我们还发现S6 K2的活性是通过与DNA的直接相互作用以剂量依赖的方式诱导的。此外，AT-钩基序的突变研究揭示了其在调节S6 K2促生存信号传导中的重要性。为了进一步推进这些原始的发现，我们建议研究S6 K2-DNA相互作用的分子机制，并采用一系列生物物理，生物化学和细胞的方法，S6 K2激活DNA结合后的替代模型。我们的努力也将集中在探索S6 K2和转录因子之间的相互作用的合作模式，靶向启动子和增强子区域的一组特定的基因，涉及S6 K2介导的细胞过程的调节。将采用哺乳动物细胞模型和裸鼠异种移植研究来研究S6 K2 AT-钩基序在转录、核糖体生物合成、细胞存活和生长调节中的作用。预计这项拟议的研究将更深入地了解S6 K2的核功能及其在转录调节中的意义。S6 K2-DNA相互作用的独特特征不仅具有学术意义，而且还可能提供适用于药物开发的新见解。

项目成果

Regulation of the CoA Biosynthetic Complex Assembly in Mammalian Cells.

• DOI: 10.3390/ijms22031131
• 发表时间: 2021-01-24
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Baković J;López Martínez D;Nikolaou S;Yu BYK;Tossounian MA;Tsuchiya Y;Thrasivoulou C;Filonenko V;Gout I
• 通讯作者: Gout I

Redox Regulation of the Quorum-sensing Transcription Factor AgrA by Coenzyme A.

• DOI: 10.3390/antiox10060841
• 发表时间: 2021-05-25
• 期刊: Antioxidants (Basel, Switzerland)
• 作者: Baković J;Yu BYK;Silva D;Baczynska M;Peak-Chew SY;Switzer A;Burchell L;Wigneshweraraj S;Vandanashree M;Gopal B;Filonenko V;Skehel M;Gout I
• 通讯作者: Gout I

Three-dimensional cancer cell culture in high-yield multiscale scaffolds by shear spinning

通过剪切旋转在高产多尺度支架中进行三维癌细胞培养

• DOI: 10.1002/btpr.2750
• 发表时间: 2018
• 期刊: Biotechnology Progress
• 影响因子: 2.9
• 作者: Ahmed A

iPSC-derived neuronal models of PANK2-associated neurodegeneration reveal mitochondrial dysfunction contributing to early disease.

• DOI: 10.1371/journal.pone.0184104
• 发表时间: 2017
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Arber C;Angelova PR;Wiethoff S;Tsuchiya Y;Mazzacuva F;Preza E;Bhatia KP;Mills K;Gout I;Abramov AY;Hardy J;Duce JA;Houlden H;Wray S
• 通讯作者: Wray S