CAREER: Arabidopsis Argonaute10-protein Interactome

智力优势：精氨酸(AGO)蛋白是RNA介导的基因沉默的关键效应者，通过与miRNAs结合在转录或转录后水平上抑制目标基因的表达。虽然agos以前被认为只抑制miRNA靶基因，但Pi的研究小组最近发现，拟南芥AGO10特异性地隔离单个miRNA类别miR166来积极调节他们的靶基因。这种调节的机制尚不清楚，但它可能涉及与AGO10相互作用的蛋白质。初步研究确定了AGO10的几个新的辅因子，包括两个丝氨酸和精氨酸富集型剪接因子(SRSFs)和一个AGO10相互作用的外切核酸酶(AGIER1)等。这些新发现将RNA沉默与许多以前不为人知的生物事件联系在一起，例如mRNA剪接和AGO通道的miRNA衰退。为了研究这些新的AGO10调控模式，我们将追求两个具体目标：1)研究AGO10与SRSFs之间的生化相互作用及其生物学后果；2)研究AGIER1如何影响AGO10-miR166的功能。结果将确定AGO10/miR166功能是如何调节的，并将通过miRNA功能揭示基因调控网络中新的复杂性层次。更广泛的影响：这项研究为本科生和研究生在广泛的领域提供了独特的高级培训机会。通过两项战略将研究和教育目标纳入这份职业计划：1)招聘和培训女性和少数民族本科生进行生化和遗传领域的研究生学习；2)在课堂和研究环境中实现RNA生物学研究生培训的现代化和提高质量。这些努力将有助于提供一支富有创造力和高技能的劳动力队伍，以支持美国经济。

Intellectual Merit: Argonaute (AGO) proteins are key effectors of RNA-mediated gene silencing and act by binding to miRNAs to repress the expression of target genes at transcriptional or posttranscriptional levels. Whereas AGOs were previously thought to exclusively repress miRNA target genes, the PI's group recently discovered that Arabidopsis AGO10 specifically sequesters a single miRNA class, miR166 to positively regulate their targets. The mechanism of this regulation is unclear, but it may involve proteins that interact with AGO10. Preliminary studies identified several novel cofactors of AGO10, including two serine and arginine enriched splicing factors(SRSFs) and an AGO10-Interacting exoribonuclease (AGIER1), among others. These new findings interconnect RNA silencing with many previously unappreciated biological events such as mRNA splicing and AGO-channeled miRNA decay. To investigate these novel modes of AGO10 regulation, two specific objectives will be pursued: 1) Investigate the biochemical interaction between AGO10 and the SRSFs and its biological consequences; 2) Investigate how AGIER1 affects AGO10-miR166 function. Results will establish how AGO10/miR166 function is modulated and will reveal new layers of complexity in gene regulatory networks through miRNA function. Broader Impacts: The research provides unique and advanced training opportunities for undergraduates, and graduate students in broad areas. Research and educational objectives are integrated in this CAREER proposal through two strategies: 1) to recruit and train female and minority undergraduates for graduate studies in biochemical and genetic areas; and 2) to modernize and improve the quality of graduate training in RNA biology in classroom and research settings. These efforts will contribute to providing a creative and highly skilled workforce to support the U.S. economy.