BIOCHEMISTRY OF REGULATED PRE-MRNA SPLICING/DROSOPHILA

调控前 mRNA 剪接的生物化学/果蝇

• 批准号: 7990617
• 负责人: DONALD C RIO
• 金额: $ 27.2万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-17 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7990617
• 关键词: Affect; Alternative Splicing; Biochemical; Biochemistry; Development; Disease; Drosophila genus; Drosophila melanogaster; Excision; Gene Expression; Gene Mutation; Genomics; Grant; Human; Informatin; Introns; Mus; Nuclear; Pattern; Phosphorus; Play; Process; Protein Structure Initiative; Proteins; Proteomics; RNA Binding; RNA Processing; RNA Splicing; RNA-Binding Proteins; Regulation; Research; Role; Squid; Structure; Therapeutic Intervention; genome-wide; mRNA Precursor

Alternative pre-mRNA splicing is a common mechanism for regulating gene expression in metazoans. Indeed, RNA processing is a conduit by which genomic information is transferred to proteomic information. In fact, it is now recognized that 40-50% of the known human and mouse disease gene mutations affect the splicing process. Thus, understanding how introns are recognized and how patterns of alternative splicing are set up may allow therapeutic intervention. The overall objective of the research proposed in this grant is to elucidate biochemical mechanisms used to regulate splicing of messenger RNA precursors (pre-mRNA) during development in the fruit fly,Drosophila melanogaster. Our efforts will be focused on understanding two examples of alternative splicing in detail and, more generally, the role that RNA binding proteins play in controlling splicing patterns. In order to accomplish our overall objectives, we will: 1. Purify and analyze nuclear pre-mRNPs that contain the Drosophila hnRNP proteins, PSI, hrp48, hrp40, hrp38 and hrp36. 2. Investigate the splicing regulation of the P element and squid/hrp40 pre-mRNAs using biochemical studies. 3. Analyze the genome-wide effects on alternative pre-mRNA splicing patterns after removal of specific RNA binding splicing factors using Drosophila splice junction microarrays. 4. Analyze the structure and function of PSI, Drosophila U2AF and U2AF-related proteins.

选择性的前mRNA剪接是调节后生动物基因表达的一种常见机制。 的确，RNA处理是基因组信息传递到蛋白质组信息的管道。 事实上，现在人们已经认识到，已知的人类和老鼠疾病基因突变中有40%-50%会影响 拼接过程。因此，了解内含子是如何识别的，以及选择性剪接模式是如何 可以允许治疗性干预。这项拨款建议的研究的总体目标是 阐明调控信使RNA前体剪接的生化机制 果蝇在发育过程中，黑腹果蝇。我们的努力将集中在理解 详细介绍了两个选择性剪接的例子，以及更一般的RNA结合蛋白在其中所起的作用 控制拼接模式。为了实现我们的总体目标，我们将： 1.提纯和分析含有果蝇hnRNP蛋白、PSI、hrp48、hrp40、 Hrp38和hrp36。 2.用生化方法研究P元件与Squid/hrp40前-mRNAs的剪接规律 学习。 3.分析去除特定基因后全基因组范围内对不同的前-mRNA剪接模式的影响 利用果蝇剪接连接微阵列的RNA结合剪接因子。 4.分析PSI、果蝇U2AF和U2AF相关蛋白的结构和功能。

项目成果

Genome-wide identification of alternative splice forms down-regulated by nonsense-mediated mRNA decay in Drosophila.

• DOI: 10.1371/journal.pgen.1000525
• 发表时间: 2009-06
• 期刊: PLoS genetics
• 影响因子: 4.5
• 作者: Hansen KD;Lareau LF;Blanchette M;Green RE;Meng Q;Rehwinkel J;Gallusser FL;Izaurralde E;Rio DC;Dudoit S;Brenner SE
• 通讯作者: Brenner SE

Evolution of a tissue-specific splicing network.

• DOI: 10.1101/gad.2009011
• 发表时间: 2011-03
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: J. Matthew Taliaferro;Nehemiah S. Alvarez;Richard E. Green;M. Blanchette;D. C. Rio

A segmental genomic duplication generates a functional intron.

• DOI: 10.1038/ncomms1461
• 发表时间: 2011-08-30
• 期刊: Nature communications
• 影响因子: 16.6

The Drosophila splicing factor PSI is phosphorylated by casein kinase II and tousled-like kinase.

果蝇剪接因子 PSI 被酪蛋白激酶 II 和蓬乱样激酶磷酸化。

• DOI: 10.1371/journal.pone.0056401
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Taliaferro,JMatthew;Marwha,Dhruv;Aspden,JulieL;Mavrici,Daniela;Cheng,NathalieE;Kohlstaedt,LoriA;Rio,DonaldC
• 通讯作者: Rio,DonaldC

A green fluorescent protein solubility screen in E. coli reveals domain boundaries of the GTP-binding domain in the P element transposase.

大肠杆菌中的绿色荧光蛋白溶解度筛选揭示了 P 元件转座酶中 GTP 结合结构域的结构域边界。

• DOI: 10.1002/pro.499
• 发表时间: 2010
• 期刊: Protein science : a publication of the Protein Society
• 作者: Sabogal,Alex;Rio,DonaldC
• 通讯作者: Rio,DonaldC