Structure and Function of Nucleic Acids

描述（由申请人提供）：长期目标是能够从其序列中正确预测（1）RNA如何折叠成其生物学功能形式，（2）其结构如何影响其功能，以及（3）其折叠速度和对环境中蛋白质和其他配体的反应速度。RNA分子在基因表达的各个方面都至关重要。信使RNA将遗传信息从DNA转移到蛋白质;在RNA病毒中，RNA既是遗传信息，也是蛋白质合成的信使。非编码RNA，包括microRNA和小干扰RNA，调节DNA转录为RNA和RNA翻译为蛋白质。信使RNA的加工和控制错误与人类疾病有关，包括神经退行性疾病，如Lou Gehrig病。RNA作为信使RNA的抑制剂被研究，以沉默特定的基因，并治愈疾病。RNA--在人类、病毒和细菌中--也是预防或治疗人类疾病的药物的突出靶点。了解RNA的三维结构、稳定性和相互转化率对于理解RNA的所有不同生物功能至关重要。 为了获得这些知识，应用了新发现的单分子方法：激光镊子和荧光共振能量转移（FRET）。RNA分子在实验室中通过RNA聚合酶从DNA模板合成。每个RNA分子上都附有一个特异性标记，以便对其进行监测和表征。激光镊子：将一个微米大小的珠子连接到单个RNA分子的两端;一个珠子放在微量移液管中，另一个放在激光陷阱中。当RNA分子移动时，或者当它与其他RNA或蛋白质相互作用时改变形状，珠子就会移动。它们的运动可以在显微镜下看到，从而评估RNA的行为。FRET：荧光染料被放置在RNA或与之相互作用的分子上。来自染料的荧光信号对染料之间的距离非常敏感，因此对RNA的运动和形状非常敏感。这些数据提供了RNA的热力学稳定性及其解折叠和重折叠的速率。确定了解开、转录和翻译RNA的蛋白质以及抑制这些过程的药物的作用。这些信息将有助于我们更好地理解RNA的结构、稳定性和动力学。它将有助于理解RNA的功能，并控制RNA在人类疾病中的作用。

DESCRIPTION (provided by applicant): The long term goal is to be able to correctly predict from its sequence (1) how an RNA folds into its biologically functional form, (2) how its structure affects its function, and (3) how fast it folds and responds to proteins and other ligands in its environment. RNA molecules are crucial in all aspects of gene expression. Messenger RNAs transfer the genetic information from DNA to proteins; in RNA viruses the RNA is both the genetic information, and the messenger for protein synthesis. Non-coding RNAs, including microRNAs and small interfering RNAs, regulate transcription of DNA to RNA and translation of RNA to protein. Errors in processing and control of messenger RNAs are linked to human diseases, including neurodegenerative diseases such as Lou Gehrig's disease. RNAs are being investigated as inhibitors of messenger RNAs to silence specific genes, and to cure diseases. RNAs - in humans, viruses, and bacteria - are also outstanding targets for drugs to prevent or cure human diseases. Knowledge of the three-dimensional structures of RNAs, their stabilities, and their rates of interconversion is crucial to understanding all the different biological functions of RNA. In order to obtain this knowledge, newly discovered single-molecule methods are applied: Laser Tweezers and Fluorescence Resonance Energy Transfer (FRET). RNA molecules are synthesized in the laboratory by RNA polymerase from a DNA template. A specific label is attached to each RNA molecule so that it can be monitored and characterized. Laser Tweezers: A micron-sized bead is attached to each end of a single RNA molecule; one bead is held in a micropipette, the other in a laser trap. As the RNA molecule moves, or changes shape as it interacts with other RNAs or proteins, the beads move. Their motion can be seen in a microscope and thus the behavior of the RNA is assessed. FRET: Fluorescent dyes are placed on the RNA or on the molecules it interacts with. The fluorescent signals from the dyes are exquisitely sensitive to the distance between the dyes, and thus on the motion and shape of the RNA. These data provide the thermodynamic stability of the RNA, and its rates of unfolding and refolding. The effects of proteins that unwind, transcribe, and translate the RNA, as well as drugs that inhibit these processes are determined. This information will lead to improved understanding of RNA structure, stability, and dynamics. It will help in understanding RNA function, and in controlling the role of RNA in human diseases.