DETERMINANTS OF SPECIFICITY IN PROTEIN-RNA INTERACTIONS

本项目旨在阐明核糖核酸的作用机制。 RNA结合蛋白家族(RRM家族)的识别 一个常见的序列主题。RRM家族的新成员将是 用简并核酸探针法和抗体反应性分离。 将使用分子遗传学和生物物理方法来阐明氨基酸。 RRM家族参与RNA结合特异性的酸性决定因素 在转录(La和Ro)和RNA加工中起作用的成员 (U1和U2SnRNP蛋白)。一个目标将集中在一种新的神经元上-- 我们已经发现了一种特殊的RRM蛋白，称为Hel-N1，可以影响细胞 在培养细胞中表达时的增殖和分化。我们的 有数据表明，Hel-N1可以与RNA中的不稳定序列结合 癌蛋白和淋巴因子信息的非编码区，包括c- MYC、c-fos、GM-CSF。我们建议研究Hel-N1和其他病毒的作用 RNA结合蛋白在RNA稳定性、生长控制和细胞中的作用 转型。因此，这些研究将解决基本问题 RNA识别和结合在多个水平上发挥作用的机制 核酸代谢和调节核酸-蛋白质的 互动。

This project is directed at the elucidation of mechanisms of RNA recognition by a family of RNA-binding proteins (RRM family) which have a common sequence motif. Novel members of the RRM family will be isolated using degenerate nucleic acid probing and antibody reactivity. Molecular genetic and biophysical methods will be used to elucidate amino acid determinants involved in RNA binding specificity by RRM family members which function in transcription (La and Ro) and RNA processing (U1 and U2 snRNP proteins). One aim will focus on a novel neuron- specific RRM protein, termed Hel-N1, which we have found can affect cell proliferation and differentiation when expressed in cultured cells. Our data indicate that Hel-N1 can bind to RNA "instability sequences" in the noncoding regions of oncoprotein and lymphokine messages, including c- myc, c-fos and GM-CSF. We propose to study the role of Hel-N1 and other RNA binding proteins in RNA stability, growth control and cellular transformation. Therefore, these studies will address fundamental mechanisms of RNA recognition and binding which function at many levels of nucleic acid metabolism and which regulate nucleic acid-protein interactions.