NSF-BSF: Deciphering the factors that govern the temperature-dependence of ADAR reactivity and dsRNA substrate recognition

RNA编辑是在包括人类在内的许多生物中发现的一种过程，能够拦截遗传信息并改变编码信息。这使得一个基因可以产生一系列不同的蛋白质。不同生物体通过编辑来调节RNA信息内容的能力可能对生物体如何应对它们遇到的压力（例如温度变化、暴露于有害物质或病原体等）产生重大影响。这项合作涉及美国的两个实验室和以色列的两个实验室，将研究来自不同生物体的RNA编辑酶ADARs（作用于RNA的腺苷脱氨酶）。这些努力将促进我们对编辑如何被生物体在其独特环境中茁壮成长的理解。该项目为分子生物学、核酸化学、酶学和结构生物学等学科的学生提供了一个独特的跨学科培训机会。每个实验室将贡献不同的学科努力，以深入了解ADARs的RNA编辑作为体温的功能，这在气候变化中是至关重要的。该团队将通过向包括RNA编辑科学界，研究生，本科生，K-12学生和公众在内的感兴趣的社区传播我们的结果，最大限度地扩大研究发现的广泛影响。拟议的新工作是NSF-BSF联合资助的；它将由彼得·比尔教授和安德鲁·费舍尔教授领导的美国团队和谢伊·本-阿罗亚教授和埃雷兹·莱瓦农教授领导的以色列团队进行。Ben-Aroya/Levanon团队已经建立了面包酵母，酿酒酵母，作为一个多功能筛选平台，以评估来自不同生物的ADARs的编辑活性。利用这个平台，他们发现鸟类ADARs是酵母中高效的腺苷到肌苷（A到I）编辑器，并提出了这是鸟类高体温的结果的假设。在这个项目中，所有四个实验室将共同努力，确定导致高编辑活性的鸟类ADARs的关键特征，并确定其分子基础。这将通过区域交换和随机诱变以及酵母生长效应的额外筛选来完成。纯化的鸟类ADAR蛋白将进行RNA结合、脱氨率测量和ADAR-RNA复合物的结构研究。最后，来自其他生物的adar，居住在不同的环境生态位，将被表征，以定义观察到的现象的普遍性。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

RNA editing is a process found in many organisms, including humans, capable of intercepting genetic messages and changing the encoded information. This allows for a range of different proteins to be made from a single gene. The ability of different organisms to modulate the information content in RNA through editing is likely to have a substantial impact on how organisms’ respond to stresses they encounter (e.g. changes in temperature, exposure to harmful substances or pathogens, etc). This collaborative effort involving two labs in the U.S. and two labs in Israel will investigate RNA editing enzymes called ADARs (adenosine deaminases that act on RNA) from different organisms. These efforts will advance our understanding of how editing can be used by an organism to thrive in its unique environment. This project provides a unique opportunity for interdisciplinary training of students spanning molecular biology, nucleic acid chemistry, enzymology, and structural biology. Each lab will contribute a different disciplinary effort to achieve in-depth understanding of RNA editing by ADARs as a function of body temperature, which is critically important in a changing climate. The team will maximize the broader impact of the research discoveries by disseminating our results to interested communities including the RNA editing scientific community, graduate students, undergraduate students, K-12 students and the general public. The proposed new work is a joint NSF-BSF grant; it will be carried out by an American team directed by Professors Peter Beal and Andrew Fisher and an Israeli effort led by Professors Shay Ben-Aroya and Erez Levanon. The Ben-Aroya/Levanon team have established the baker’s yeast, S. cerevisiae, as a versatile screening platform to evaluate the editing activity of ADARs from different organisms. Using this platform, they identified bird ADARs as highly efficient adenosine-to-inosine (A to I) editors in yeast and have advanced the hypothesis that this is a consequence of the high body temperature of birds. In this project, all four labs will work together to define the critical features of bird ADARs that lead to the high editing activity and define its molecular basis. This will be accomplished using domain swapping and random mutagenesis coupled with additional screens for growth effects in yeast. Purified bird ADAR proteins will be subjected to RNA binding, deamination rate measurements and structural studies of ADAR-RNA complexes. Finally, ADARs from other organisms, that inhabit different environmental niches, will be characterized to define the generality of the observed phenomenon.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.