HTS for Discovery of Inhibitors of STAR Protein-RNA complexes

描述(由申请人提供)：这项提案概述了一项计划，该计划旨在开发一种有效的高通量筛选，用于两种明星蛋白：GLD-1和Quking形成RNA-蛋白质复合体的抑制剂。最终目标将是配置这些分析，以便提交给MLSCN进行大规模筛查工作。GLD-1是线虫生殖系发育的调节因子，Quking是一种小鼠蛋白，调节发育中的神经系统的髓鞘形成。STAR蛋白家族是一类重要的RNA结合蛋白，还包括Sam68和SLM蛋白，它们参与了广泛的过程，并与某些类型的癌症有关。将开发一种荧光偏振分析方法，用于监测荧光标记的RNA与蛋白质结合时的偏振变化。小分子竞争者将通过筛选极化降低来识别。这些经过验证的抑制剂将是在线虫和小鼠身上进行功能研究的强大工具，一旦确定了合适的RNA靶标，该检测将很容易扩展到Sam68的抑制剂。

DESCRIPTION (provided by applicant): This proposal outlines a plan to develop a validated high throughput screen for inhibitors of RNA-protein complex formation by two STAR proteins: GLD-1 and Quaking. The ultimate goal will be to configure these assays for submission to the MLSCN for a large scale screening effort. GLD-1 is a regulatory of germline development in C. elegans, and Quaking is a mouse protein that regulates myelination in the developing nervous system. The STAR protein family is an important class of RNA binding proteins that also includes Sam68 and the SLM proteins that are implicated in a wide variety of processes, and is linked to certain types of cancer. A fluorescence polarization assay will be developed that monitors the change in polarization of a fluorescently labeled RNA upon binding to the protein. Small molecule competitors will be identified by screening for decreases in polarization. Such validated inhibitors would be powerful tools for functional studies in C. elegans and mice, and the assay should be readily extended to inhibitors of Sam68 once suitable RNA targets have been identified.