Amino Acid Regulation of Alternative Splicing

选择性剪接的氨基酸调控

• 批准号: 8705504
• 负责人: MICHAEL S. KILBERG
• 金额: $ 31.58万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8705504
• 关键词: Address; Alternative Splicing; Amino Acids; Anti-Inflammatory Agents; Anti-inflammatory; Area; Arginine; Aspartate-Ammonia Ligase; Biological Assay; CCAAT-Enhancer-Binding Proteins; Cell Culture Techniques; Cell physiology; Cells; Cellular Stress; Complementary DNA; Cultured Cells; Data; Diabetes Mellitus; Diet; Dietary Proteins; Disease; Dominant-Negative Mutation; EMSA; Embryo; Essential Amino Acids; Exhibits; Exons; Feedback; Fetal Development; Fibroblasts; Gene Targeting; Genes; Genetic Transcription; Health; Histidinol; Human; Hybrids; Individual; Inflammation; Inflammatory; Informatin; Investigation; Kinetics; Knock-out; Knowledge; Length; Leucine Zippers; Longevity; MAP Kinase Gene; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Messenger RNA; Microarray Analysis; Monitor; Mus; Nutrient; Pathway interactions; Phosphorylation; Play; Protein Array; Protein Isoforms; Protein Microarray Assay; Protein Microchips; Protein-Restricted Diet; Proteins; RNA; RNA Splicing; Recruitment Activity; Regulation; Reporter Genes; Reporting; Research; Response Elements; Role; Serine; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Small Interfering RNA; Testing; Tetanus Helper Peptide; Transgenic Mice; Tumor Promoters; Yeasts; acid stress; acronyms; activating transcription factor; chromatin immunoprecipitation; deprivation; feeding; gene repression; hepatoma cell; human ARMET protein; human FRAP1 protein; hypertensive heart disease; in vivo; insight; knock-down; mRNA Precursor; novel; nutrition; programs; protein intake; protein protein interaction; research study; response; transcription factor; tumor

DESCRIPTION (provided by applicant): Limiting dietary protein intake results in amino acid deficiency within cells and activates several signal transduction pathways collectively called the amino acid response (AAR). A number of genes have been identified that are transcriptionally-activated by the AAR, including the bZIP transcription factor ATF3, for which cellular stress induces multiple isoforms by pre-mRNA alternative splicing. Two of these isoforms, full- length ATF3 (ATF3-FL) and a form with a truncated leucine zipper, ATF3?Zip3, are induced in expression by low protein diet in vivo or by amino acid deprivation of cultured cells. These two isoforms exhibit opposing action on the AAR target gene encoding asparagine synthetase (ASNS); exogenous ATF3-FL expression causes transcriptional repression of the amino acid-dependent induction of ASNS, whereas ATF3?Zip3 further enhances the induction. How the cellular amino acid content signals to and controls pre-mRNA alternative splicing has not been investigated. In fact, the study of the regulation of alternative splicing by macro-nutrients represents an entirely new area of investigation in the splicing field. The hypothesis is that ATF3 isoforms have opposing actions within the cellular response to protein/amino acid stress and that the individual isoforms interact with activity-modifying proteins and/or transcriptional co- regulators that support these opposing activities. To address this global hypothesis, three sub- hypotheses will be tested. Hypothesis I: There are differences in the synthesis and functional activities of specific ATF3 isoforms induced by dietary low protein in mice and amino acid deprivation of cultured cells. The proposed research will investigate the kinetics of synthesis for ATF3-FL and ATF3?Zip3 and the functional consequences of each isoform will be addressed by RNA and protein microarray analysis in transgenic mice expressing either ATF3-FL or ATF3?Zip3 individually. Hypothesis II: Amino acid-dependent signaling pathways regulate the alternative splicing of ATF3 during the AAR. These studies will determine the signaling pathway responsible for sensing and transducing the amino acid deficiency signal to the proteins that regulate exon choice during alternative splicing. Hypothesis III: Protein-protein interactions of individual ATF3 isoforms modulate their action on AAR target genes. ATF3-interacting proteins will be identified and their role in the AAR determined. Collectively, the proposed studies will provide novel information and address significant gaps in our knowledge of ATF3 alternative splicing and ATF3 isoform function. The insight gained from these studies will impact the fields of: 1) macro-nutrient control of pre- mRNA alternative splicing; 2) amino acid-dependent control of transcription; and 3) ATF3 function in nutrition and disease.

描述（由申请人提供）：限制膳食蛋白质摄入导致细胞内氨基酸缺乏，并激活统称为氨基酸反应（AAR）的几种信号转导途径。已经鉴定了许多被AAR转录激活的基因，包括bZIP转录因子ATF 3，细胞应激通过前mRNA选择性剪接诱导其多种亚型。其中两种亚型，全长ATF 3（ATF 3-FL）和一种具有截短亮氨酸拉链的形式，ATF 3？Zip 3在体内通过低蛋白饮食或通过培养细胞的氨基酸剥夺诱导表达。这两种亚型表现出相反的行动上的AAR靶基因编码天冬酰胺合成酶（ASNS）;外源性ATF 3-FL表达导致转录抑制的氨基酸依赖性诱导ASNS，而ATF 3？Zip 3进一步增强了诱导。细胞氨基酸含量如何向前体mRNA可变剪接发出信号并控制前体mRNA可变剪接尚未研究。事实上，研究宏量营养素对可变剪接的调节是剪接领域一个全新的研究领域。假设是ATF 3同种型在对蛋白质/氨基酸应激的细胞应答中具有相反的作用，并且单个同种型与支持这些相反活性的活性修饰蛋白和/或转录共调节因子相互作用。为了解决这一总体假设，将检验三个子假设。假设一：低蛋白饮食和培养细胞氨基酸剥夺诱导的ATF 3特异性异构体的合成和功能活性存在差异。拟议的研究将调查ATF 3-FL和ATF 3？Zip 3和每个亚型的功能后果将通过在转基因小鼠中表达ATF 3-FL或ATF 3？Zip 3个人假设二：AAR过程中，ATF 3的选择性剪接受氨基酸依赖性信号通路调控。这些研究将确定负责感知和转导氨基酸缺陷信号的信号传导途径，以调节选择性剪接过程中外显子选择的蛋白质。假设III：单个ATF 3亚型的蛋白质-蛋白质相互作用调节它们对AAR靶基因的作用。ATF 3相互作用的蛋白质将被确定，并确定其在AAR中的作用。总的来说，拟议的研究将提供新的信息，并解决我们对ATF 3选择性剪接和ATF 3亚型功能的认识存在的重大差距。从这些研究中获得的见解将影响以下领域：1）前mRNA选择性剪接的宏观营养控制; 2）转录的氨基酸依赖性控制;以及3）营养和疾病中的ATF 3功能。

项目成果

Activation of the amino acid response modulates lineage specification during differentiation of murine embryonic stem cells.

氨基酸反应的激活调节小鼠胚胎干细胞分化过程中的谱系规范。

• DOI: 10.1152/ajpendo.00136.2013
• 发表时间: 2013
• 期刊: American journal of physiology. Endocrinology and metabolism
• 作者: Shan,Jixiu;Hamazaki,Takashi;Tang,TiffanyA;Terada,Naohiro;Kilberg,MichaelS
• 通讯作者: Kilberg,MichaelS

Asparagine Synthetase Deficiency causes reduced proliferation of cells under conditions of limited asparagine.

• DOI: 10.1016/j.ymgme.2015.08.007
• 发表时间: 2015-11
• 期刊: MOLECULAR GENETICS AND METABOLISM
• 影响因子: 3.8
• 作者: Palmer, Elizabeth Emma;Hayner, Jaclyn;Sachdev, Rani;Cardamone, Michael;Kandula, Tejaswi;Morris, Paula;Dias, Kerith-Rae;Tao, Jiang;Miller, David;Zhu, Ying;Macintosh, Rebecca;Dinger, Marcel E.;Cowley, Mark J.;Buckley, Michael F.;Roscioli, Tony;Bye, Ann;Kilberg, Michael S.;Kirk, Edwin P.
• 通讯作者: Kirk, Edwin P.

Tumor suppressor BTG1 promotes PRMT1-mediated ATF4 function in response to cellular stress.

• DOI: 10.18632/oncotarget.6519
• 发表时间: 2016-01-19
• 期刊: Oncotarget
• 作者: Yuniati L;van der Meer LT;Tijchon E;van Ingen Schenau D;van Emst L;Levers M;Palit SA;Rodenbach C;Poelmans G;Hoogerbrugge PM;Shan J;Kilberg MS;Scheijen B;van Leeuwen FN
• 通讯作者: van Leeuwen FN