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Request fund to purchase equipment (the BioComp gradient primer) to supplement R15 GM139107-01

申请资金购买设备（BioComp梯度引物）以补充R15 GM139107-01

基本信息

批准号：
10582006
负责人：
Ke Zhang Reid
金额：
$ 3.93万
依托单位：
WAKE FOREST UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-31
项目状态：
已结题

项目摘要

Project Summary (Abstract): We are applying for a supplement fund to purchase a BioComp gradient primer for fractionating density gradients. This equipment is essential for the success of the parent award as several proposed experiments, including polysome profiling, ribosome profiling, and Translating Ribosome Affinity Purification (TRAP), rely on this equipment to receive consistent and reproducible results. We desperately need this equipment to continue our investigation following equipment malfunction of our current system that cannot be repaired (system age and non-availability of parts). Additionally, no other laboratory at our institution has similar equipment we could share. Our talented undergraduate and graduate students will use this equipment to address an exciting but disputed question: does the functional specialization of ribosomes exist to support selective translation? We focus on two paralogs of predicted large ribosomal subunit 27 (Rpl27) found in fission yeast. Although these paralogs have 90% amino acid similarity and are highly conserved through much of eukaryotic evolution, their mutants demonstrate distinct phenotypes. Our preliminary data suggest that they might be part of heterogeneous ribosomes that regulate the translation of different mRNA transcripts. Dissecting the functional differences between Rpl27 paralogs may provide valuable insight into how ribosome protein paralogs potentially play a role in the heterogeneity of ribosomes, inducing a conformational change that ultimately results in altered activity and preference of mRNA transcripts accessible for translation.

项目摘要（Abstract）：我们正在申请补充基金来购买BioComp梯度引物，分级密度梯度。该设备对于父母的成功至关重要作为几个拟议的实验，包括多核糖体分析，核糖体分析和翻译核糖体亲和纯化（TRAP）依赖于该设备以获得一致和可重复的结果。我们迫切需要这些设备继续我们的调查后，设备故障，我们目前的系统，无法修复（系统老化和部件不可用）。此外，没有其他我们实验室有类似的设备，我们可以共享。我们才华横溢本科生和研究生将使用这种设备来解决一个令人兴奋的，但一个有争议的问题：核糖体的功能特化是否支持选择性翻译？我们集中在预测的核糖体大亚基27的两个旁系同源物（Rp127）在分裂酵母中发现。虽然这些旁系同源物有90%的氨基酸相似性在真核生物进化过程中高度保守，表现出不同的表型。我们的初步数据显示它们可能是调节不同mRNA转录本翻译的异质核糖体。剖析Rp127旁系同源物之间的功能差异可以提供有价值的信息。深入了解核糖体蛋白旁系同源物如何在细胞异质性中发挥作用，核糖体，诱导构象变化，最终导致活性改变和mRNA转录物可用于翻译的偏好。