Regulation of RNA metabolism and cell growth control by SR protein kinases

SR 蛋白激酶对 RNA 代谢和细胞生长控制的调节

• 批准号: 7196915
• 负责人: XIANG-DONG FU
• 金额: $ 31.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7196915
• 关键词: Active Sites; Address; Affect; Agar; Alternative Splicing; Animal Model; Animals; Attention; Brain; Cell Nucleus; Cell Survival; Cell model; Cells; Class; Classification; Complex; Control Animal; Coupling; Cytoplasm; Development; Disease; Embryo; Embryonic Development; Enzymes; Event; Family; Family member; Fibroblasts; Funding; G2 Phase; Gene Expression; Genetic; Genetic Transcription; Genome Stability; Goals; Heart; Individual; Interphase; Knock-out; Knockout Mice; Link; Localized; Mammalian Cell; Mammals; Mediating; Metabolism; Modeling; Mus; Nuclear Import; Nuclear Translocation; Nude Mice; Numbers; Oncogenes; Pathway interactions; Phase; Phenotype; Phosphorylation; Phosphotransferases; Play; Positioning Attribute; Post-Transcriptional Regulation; Process; Protein Dephosphorylation; Protein Kinase; Protein Sortings; Proteins; Publishing; RNA; RNA Splicing; Regulation; Research; Research Personnel; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Soft Agar Assay; Structure; System; Testing; Thymus Gland; Tissues; Transgenic Mice; Translations; Tumor Suppression; Tumor Suppressor Genes; Tumor Suppressor Proteins; Work; base; cell growth; design; in vivo; insight; interest; mRNA Export; messenger ribonucleoprotein; metaplastic cell transformation; mouse model; programs; research study; response; subcutaneous; tumor; yeast two hybrid system

DESCRIPTION (provided by applicant): Post-transcriptional regulation of gene expression is critical for cell growth control and animal development. This project attacks SRPK1 and 2, 2 evolutionary conserved kinases that correspond to the major kinase activity for the SR family of splicing factors and regulators in mammalian cells. SR proteins are involved in many aspects of RNA metabolism, all of which appear to be regulated by phosphorylation. Strikingly, we recently found that mouse embryo fibroblasts genetically deleted of SRPK1 are transformed in the soft agar assay and in nude mice. Furthermore, SRPKs are localized in the cytoplasm and can be induced to translocate to the nucelus in response to specific signals. Thus, SRPKs are fundamentally important for cell growth control and the kinase system may be regulated by signaling. Building upon our expertise established in the previous funding periods of this project, long-term interest in post-transcriptional regulation of gene expression, and extensive published and unpublished findings, we propose to continue this project under 3 specific aims for the next phase of research. Aim 1 is to develop conditional knockout mice models to determine the functional requirement for both SRPK1 and 2 during mouse development and characterize the kinase knockout mouse embryo fibroblasts to provide genetic evidence that SRPKs regulate the function SR proteins in RNA metabolism in mammalian cells. Aim 2 is to address the putative tumor suppressor activity of SRPK1 by testing 3 specific hypotheses: (1) SRPK1 may regulate alternative splicing of oncogenes and tumor supressors via SR proteins; (2) SRPK1-mediated phosphorylation may act to reorganize newly exported mRNA-protein complex to regulate translation in the cytoplasm; and (3) SRPK1-mediated phosphorylation may play a critical role in maintaining genomic stability by modulating the coupling between transcription and splicing. Aim 3 is to understand how SRPKs might be regulated by signaling. A panel of SRPK-interacting proteins have been identified and many are known components of various signal transduction pathways. We design specific experiments to understand how some specific signals may be transduced via SRPKs to regulate gene expression at post- transcriptional levels. Together, the project will dissect an unprecedented cellular transformation pathway.

描述（由申请方提供）：基因表达的转录后调控对于细胞生长控制和动物发育至关重要。该项目攻击SRPK 1和2，2进化保守激酶，其对应于哺乳动物细胞中剪接因子和调节因子的SR家族的主要激酶活性。SR蛋白参与RNA代谢的许多方面，所有这些似乎都受到磷酸化的调节。引人注目的是，我们最近发现SRPK 1基因缺失的小鼠胚胎成纤维细胞在软琼脂试验和裸鼠中转化。此外，SRPKs定位于细胞质中，并且可以响应于特定信号而被诱导易位到珠心。因此，SRPKs对细胞生长控制至关重要，激酶系统可能受信号传导调节。基于我们在该项目以前的资助期内建立的专业知识，对基因表达转录后调控的长期兴趣，以及广泛的已发表和未发表的研究结果，我们建议在下一阶段的研究中根据3个具体目标继续该项目。目的1：建立SRPK 1和SRPK 2基因敲除小鼠模型，研究SRPK 1和SRPK 2在小鼠发育过程中的功能需求，并对SRPK 1和SRPK 2基因敲除小鼠胚胎成纤维细胞进行鉴定，为SRPKs调控哺乳动物细胞RNA代谢提供遗传学证据。目的二是通过验证以下3个假设来阐明SRPK 1的抑癌活性：（1）SRPK 1可能通过SR蛋白调节癌基因和抑癌基因的选择性剪接：（2）SRPK 1介导的磷酸化可能通过重组新输出的mRNA-蛋白复合物来调节胞浆中的翻译;（3）SRPK 1介导的磷酸化可能通过调节转录和剪接之间的偶联而在维持基因组稳定性中起关键作用。目的3是了解SRPKs如何通过信号转导进行调节。已经鉴定了一组SRPK相互作用蛋白，其中许多是各种信号转导途径的已知组分。我们设计了特定的实验来了解一些特定的信号是如何通过SRPKs转导来在转录后水平调节基因表达的。总之，该项目将剖析一个前所未有的细胞转化途径。