Phosphoiylation -dependent regulation of alternative splicing

选择性剪接的磷酸化依赖性调控

基本信息

批准号：
14380326
负责人：
HAGIWARA Masatoshi
金额：
$ 9.6万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14380326/
关键词：
alternative splicing Phosphorylation SR proteins SRPK Clk mRNA processing protein kinase inhibitor Clk 特異的阻害剤

项目摘要

The regulation of splice site usage provides a versatile mechanism for controlling gene expression and for the generation of proteome diversity, playing an essential role in many biological processes. The selection of splice site can be altered by numerous extracellular stimuli, including growth factors, cytokines, hormones, depolarization, osmotic shock, and UVC irradiation, through synthesis, phosphorylation, and a change in localization of serine/arginine-rich (SR) proteins. SR proteins are a family of essential factors required for constitutive splicing of pre-mRNA and play an important role in modulating alternative splicing. They are highly conserved in eukaryotes and are characterized by having one or two RNA-recognition motifs (RRMs) at the amino terminus and an RS domain at the carboxy terminus.Phosphorylation state of SR proteins appears to influence their activities in general and alternative splicing. To date, several kinases have been reported to phosphorylate SR proteins, … More including SRPK-familykinases (SRPK1 and SRPK2), Clk/Sty family kinases, which consisted of four members (Clkl/Sty and Clk2-4), DNA topoisomerase I, p34cdc2 kinase and hPRP4, Addition of purified SRPK1 to permeabilized cells, or overexpression of SRPK1, 2 or one of CIk family members in transfected cells result in an apparent disassembly of the nuclear speckles.The kinase inhibitor DRB (5,6-dichloro-1-β-D-ribo-furanosylbenzimidazole), which was previously shown to inhibit casein kinase II, inhibited Clk2 and induced the redistribution of SR proteins. Recently, through extensive screening of a chemical library, a specific inhibitor of Clkl/Sty and Clk4 was found and named as TG003. Clk 1/Sty-dependent splicing and disassembly of nuclear speckles were suppressed by TG003. Moreover, administration of TG003 rescued the embryonic defects induced by excessive Clk/Sty activity in Xen opus. As the importance of alternative splicing has been illustrated by the increasing number of human diseases attributed to missplicing events, TGOO3 will be applicable for the therapeutic manipulation of.abnormal splicing induced by activated Clk/Sty.Considering the therapeutic application of inhibitors of SR protein kinases, prevention of virus is one of hopeful fields, because splicing is a crucial step for virus multiplication. In the case of HIV, 8 acceptor sites are used in competition to produce the vif, vpu, vpr, nef, env, tat and rev mRNAs, and the usage of these splicing sites are regulated by SR proteins and hnRNPs. In addition, Akusjarvi group reported that SR proteins purified from late adenovirus-infected cells are inactivated as splicing enhancer or splicing repressor proteins by virus-induced dephosphorylation. The anti-virus effects of inhibitors of SR protein kinases are now under investigation in our laboratory. Less

剪接部位使用的调节提供了一种多功能机制，可控制基因表达和产生蛋白质组多样性，在许多生物过程中起着至关重要的作用。剪接位点的选择可以通过许多细胞外刺激来改变，包括生长因子，细胞因子，激素，沉积，渗透性休克和UVC辐射，通过合成，磷酸化以及丝氨酸/精氨酸（SR）富含丝氨酸/精氨酸（SR）蛋白质的定位变化。 SR蛋白是原始剪接前MRNA所需的基本因素家族，并且在调节替代剪接方面起着重要作用。它们在真核生物中是高度保守的，其特征是在氨基末端具有一个或两个RNA - 识别基序（RRMS），而在羧基末端的RS结构域。SR蛋白的磷酸化状态似乎会影响其活性在一般和替代剪接中。迄今为止，据报道，已经进行了几种激酶来磷酸化的SR蛋白，其中包括包括SRPK-氨基氨基氨基氨基氨基酶酶（SRPK1和SRPK2），CLK/STY家族激酶，其中四个成员（CLKL/STY和CLK2-4）组成，DNA TOPOISomerase I，P34CDC2 KINASE和HPRP4酶供DNA Toperase I，prppyls purpp4，srps purp4，转染细胞中SRPK1、2或一个CIK家族成员的细胞或过表达导致核激酶抑制剂DRB（5,6-二氯1-β-β-d- ribo-呋喃糖基苄咪唑嗪）的明显拆卸，以前显示出抑制病例激酶II的细胞，并抑制了clkk2和SRENIB的clibib。最近，通过对化学库进行广泛的筛选，发现了Clkl/sty和Clk4的特定抑制剂，并将其命名为TG003。 TG003抑制了CLK 1/样式依赖性的剪接和拆卸核斑点。此外，TG003的给药反应了Xen Opus中过多的CLK/sty活性引起的胚胎缺陷。由于替代剪接的重要性已经通过归因于错过事件的人类疾病的越来越多，因此TGOO3适用于对激活的clk/stys诱导的毫无疑问的治疗操作。考虑sr蛋白基因酶的抑制作用是一种预期的sr蛋白基因疗法，以供激活的clk/stys.contin.stements provential sr pertential IS tofulcul in Vireential IS aftucture IS provuture Is to faculiture Is faculituce for for for for five。病毒繁殖。在艾滋病毒的情况下，在竞争中使用了8个受体位点来生产VIF，VPU，VPR，NEF，ENV，TAT，TAT和REV mRNA，并且这些剪接位点的使用受SR蛋白和HNRNP的调节。此外，Akusjarvi组报告说，从腺病毒后期感染的细胞中纯化SR蛋白被病毒诱导的去磷酸化的剪接增强子或剪接反射蛋白灭活。 SR蛋白激酶抑制剂的抗病毒作用现在正在我们的实验室中进行研究。较少的