Insulin Signaling Pathways Regulating PKCBeta Splicing

调节 PKCβ 剪接的胰岛素信号通路

基本信息

批准号：
8038449
负责人：
DENISE Ratzlaff COOPER
金额：
$ 27.79万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-08-15 至 2013-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8038449
关键词：
1-Phosphatidylinositol 3-Kinase 3&apos Splice Site Abbreviations Actins Address Alternative Splicing Antisense Oligonucleotides Area Arginine Binding Binding Proteins Binding Sites Biological Assay C-terminal Cell Extracts Cell Nucleus Cell physiology Cells Cessation of life Code Complementary DNA Complex DNA Polymerase II DNA Sequence Rearrangement Data Diabetes Mellitus Disease Documentation Elements Embryo Endocrine system Enhancers Esters Event Evolution Excision Exercise Exons F-Actin Family Fatty acid glycerol esters Fibroblasts Fibronectins Gene Expression Gene Targeting Genes Genetic Transcription Glucose Glucosephosphate Dehydrogenase Goals Health Healthcare Hela Cells Hormonal Hormones Human Human Genome IGF1 gene In Vitro Insulin Insulin Receptor Insulin Resistance Insulin Signaling Pathway Introns JAK2 gene Ligands Liver Mass Spectrum Analysis Mediating Messenger RNA Metabolic Modeling Molecular Monitor Muscle Muscle Cells Mutate Nature Nuclear Nuclear Extract Pathway interactions Phosphatidylinositols Phosphoric Monoester Hydrolases Phosphorylation Phosphorylation Site Phosphotransferases Play Prevalence Principal Investigator Process Protein Binding Protein Dephosphorylation Protein Isoforms Protein Kinase Protein Kinase C Protein Splicing Protein-Serine-Threonine Kinases Proteins Proto-Oncogene Proteins c-akt Pyrimidine RNA RNA Polymerase II RNA Processing RNA Recognition Motif RNA Splicing Reaction Receptor Protein-Tyrosine Kinases Receptor Signaling Regulation Repression Research Resistance Role SH2B gene STY kinase Serine Serum Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Site Site-Directed Mutagenesis Skeletal Muscle Small Interfering RNA Smooth Muscle Myocytes Sodium Dodecyl Sulfate-PAGE Solid Specificity Spliceosomes Starvation Stress Surface System TNF gene Techniques Text Tissues Transcript Translations U1 Small Nuclear Ribonucleoprotein Untranslated Regions Variant Western Blotting acrosome stabilizing factor cell type cis acting element design diabetic embryonic stem cell glucose uptake human ARMET protein insulin sensitivity insulin signaling mRNA Precursor member myotonic dystrophy protein kinase overexpression polypeptide protein function protein tyrosine phosphatase 1B receptor response src-Family Kinases therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Alternative splicing of pre-mRNA transcripts is a widespread means for producing polypeptide diversity from a single gene. Over 60% of human genes are expressed through alternative splicing, however, mechanisms of splicing regulation are poorly understood. This lab discovered that insulin regulates the alternative splicing of protein kinase C-(II (PKC(II) in its target tissues: muscle, fat, liver and in cells with functional insulin receptor such as aortic smooth muscle cells, embryonic fibroblasts, and HeLa cells. We identified members of the Serine/Arginine-rich (SR) family splicing proteins that bind to splicing enhancers in the pre-mRNA to regulate exon inclusion as the factors phosphorylated in response to insulin. We first studied SRp40, a splicing enhancer and identified Akt as a kinase that regulated its function via phosphorylation of residues in the arginine/serine (RS) domain. We hypothesize that Akt acts as a molecular switch in splicing regulation at several steps by also regulating other SR protein kinases such as Clk, a family of four dual function LAMMER kinases. Kinases such as Clk1 (also called Clk/Sty) and Clk2 phosphorylate SR proteins and alter their interactions in the spliceosome. Unraveling how Akt regulates Clk will add another level of regulation to insulin action. The long-term goal of the research is to determine how insulin regulates nuclear splice site selection via the activation of various kinases and splicing factors. The current aims will investigate (1) the roles of Clk1 and Clk2 phosphorylation in PKC( alternative splicing, (2) determine how SRp55 functions in PKC( splicing, and (3) identify spliceosome complexes involved in the insulin activated spliceosome and depletion of splicing factors from nuclear extracts using in vitro splicing assays to define cis-elements involved in insulin regulated splicing. The discovery that insulin regulates splicing of PKC(II, a kinase involved in insulin responses at multiple levels, indicates that there are also other target genes of this pathway that must also be spliced in a similar manner. The unique system will reveal the nature of kinase regulation, focusing on PKC(II in splicing and diabetes. PUBLIC HEALTH RELEVANCE Given the magnitude of the problems encountered with diabetes and its complications, understanding insulin action has an immense impact on healthcare since it is the sixth leading cause of disease-related death in the US. The need to define the factors contributing to diabetes onset and identify new potential therapeutic targets is a priority. The processing of pre-mRNA following insulin stimulation of its target tissues is a poorly understood area that is altered in the diabetic state. Understanding the insulin receptor signaling pathways with the goal of defining how insulin action is reflected in the nucleus of insulin responsive tissues will allow us to determine the specificity of signaling through the insulin receptor to regulate metabolic functions causing resistance to insulin action. This proposal is designed to investigate a new kinase in the insulin signaling cascade, Clk/Sty, and its nuclear substrates, SR proteins, which modify RNA processing to alter gene expression. SR proteins are altered in insulin resistance.

描述（由申请人提供）：前MRNA转录本的替代剪接是从单个基因产生多肽多样性的普遍手段。超过60％的人类基因是通过替代剪接表达的，但是，剪接调节的机制知之甚少。 This lab discovered that insulin regulates the alternative splicing of protein kinase C-(II (PKC(II) in its target tissues: muscle, fat, liver and in cells with functional insulin receptor such as aortic smooth muscle cells, embryonic fibroblasts, and HeLa cells. We identified members of the Serine/Arginine-rich (SR) family splicing proteins that bind to splicing enhancers in前MRNA调节外显子纳入胰岛素的磷酸化，我们首先研究了SRP40，这是一种剪接增强子，并确定AKT是一种激酶，通过在精液/丝氨酸（RS）域中通过SPITIC进行了SPICER的STRICTARTIAL STICALS SCHOTIEL SCORTULTAR SCTICTS SCOTCONT SCHOTIONT AKT磷酸化。蛋白质激酶，例如四个双重功能板酶的家族，例如CLK1（也称为CLK/sty）和CLK2磷酸化的SR蛋白质，并改变其在剪接中的相互作用。激酶和剪接因子。胰岛素调节胰岛素的胰岛素调节pkc的发现（ii，在多个级别的胰岛素反应中涉及的激酶）表明，该途径的其他目标基因也必须以类似的方式固定该途径。独特的系统将透露Kinase posies tabice and compoy，ii neverboy compoties emplice everys的链接。胰岛素调节PKC的剪接（ii，在多个级别上涉及胰岛素反应的激酶，表明该途径的其他靶基因也必须以类似的方式固定该途径。糖尿病及其并发症的问题对医疗保健有很大的影响，因为这是美国与疾病相关的第六个主要原因胰岛素受体信号传导途径的目的是确定胰岛素反应性组织核中如何反映胰岛素作用，这使我们能够通过胰岛素受体来确定信号传导的特异性，以调节该胰岛素的抑制作用，以调节胰岛素的新动力酶，并构成了胰岛素casce casce/clkk/clkk/clkk casce casce case/clkk/clkk/ RNA处理以改变基因表达。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The protein kinase C beta II exon confers mRNA instability in the presence of high glucose concentrations.

蛋白激酶 C beta II 外显子在高葡萄糖浓度下导致 mRNA 不稳定。

DOI：
10.1074/jbc.m206797200
发表时间：
2003
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Patel,NiketaA;Eichler,DuaneC;Chappell,DavidS;Illingworth,PhilipA;Chalfant,CharlesE;Yamamoto,Mayumi;Dean,NicholasM;Wyatt,JacquelineR;Mebert,Konrad;Watson,JamesE;Cooper,DeniseR
通讯作者：
Cooper,DeniseR

PKCdelta alternatively spliced isoforms modulate cellular apoptosis in retinoic acid-induced differentiation of human NT2 cells and mouse embryonic stem cells.

DOI：
10.3727/000000006783991890
发表时间：
2006
期刊：
Gene expression
影响因子：
0
作者：
Patel NA;Song SS;Cooper DR
通讯作者：
Cooper DR

Phosphoinositide 3-kinase mediates protein kinase C beta II mRNA destabilization in rat A10 smooth muscle cell cultures exposed to high glucose.

在暴露于高葡萄糖的大鼠 A10 平滑肌细胞培养物中，磷酸肌醇 3-激酶介导蛋白激酶 C beta II mRNA 不稳定。

DOI：
10.1016/s0003-9861(02)00208-4
发表时间：
2002
期刊：
Archives of biochemistry and biophysics
影响因子：
3.9
作者：
Patel,NiketaA;Yamamoto,Mayumi;Illingworth,Philip;Mancu,Daniel;Mebert,Konrad;Chappell,DavidS;Watson,JamesE;Cooper,DeniseR
通讯作者：
Cooper,DeniseR

Insulin regulates protein kinase CbetaII alternative splicing in multiple target tissues: development of a hormonally responsive heterologous minigene.

胰岛素调节多个靶组织中的蛋白激酶 CbetaII 选择性剪接：激素响应异源小基因的发育。

DOI：
10.1210/me.2003-0391
发表时间：
2004
期刊：
Molecular endocrinology (Baltimore, Md.)
影响因子：
0
作者：
Patel,NiketaA;Apostolatos,HerculesS;Mebert,Konrad;Chalfant,CharlesE;Watson,JamesE;Pillay,TS;Sparks,Janet;Cooper,DeniseR
通讯作者：
Cooper,DeniseR