Combinatoial CREB/ATF dimers and cellular growth control

CREB/ATF二聚体组合和细胞生长控制

• 批准号: 6608067
• 负责人: Wayne P Wahls
• 金额: $ 24.37万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6608067
• 关键词: DNA binding protein; Schizosaccharomyces pombe; X ray crystallography; binding sites; biological signal transduction; cAMP response element binding protein; cell growth regulation; dimer; gel mobility shift assay; mass spectrometry; matrix assisted laser desorption ionization; mitogen activated protein kinase; polymerase chain reaction; protein protein interaction; stress proteins; transcription factor

APPLICANT'S DESCRIPTION: Proteins of the ATF/CREB/AP-1 family are components of signal transduction pathways that monitor intracellular and extracellular conditions and transmit those signals to downstream targets. Some family members (e.g., vFos and vJun) are oncogenic. ATF/CREB/AP-1 proteins share a conserved bZIP domain that mediates both protein dimerization and sequence-specific DNA binding activity. While the bZIP domains are highly homologous to one another, some combinations of bZIP proteins form dimers, while others do not. Furthermore, different bZIP dimers have the remarkable ability to discriminate between closely related DNA binding sites. This enables a limited set of bZIP monomers to form a repertoire of heterodimers, each of which presumably regulates the expression of a specific set of genes involved in a particular biological response. An understanding of which bZIP protein dimers can form, what DNA sites they occupy, and how such events are regulated will reveal fundamental aspects in the control of cellular homeostasis and/or differentiation. The fission yeast S. pombe provides an attractive model organism for such a study. We have shown that bZIP proteins of fission yeast form combinatorial dimers in vivo and in vitro. The various dimers are under control of a MAP kinase cascade and different dimers elicit distinct effector functions. Because S. pombe has a small genome (about 6,000 genes), and the sequencing project is nearly complete, one can identify and analyze the majority of cellular bZIP proteins. Since mutants lacking bZIP proteins and signal transduction proteins are generally viable, it is also possible to reveal both upstream regulatory and downstream effector functions. A comprehensive and systematic analysis of bZIP protein dimers of fission yeast is proposed. Aim 1, To elucidate the bZIP alphabet of S. pombe: determine pairwise combinatorial associations of bZIP dimers and identify DNA sites to which they bind with high affinity. Aim 2, To reveal molecular determinants of combinatorial dimer formation and DNA binding: compare high-resolution structures of bZIP dimers alone and complexed to their DNA sites. Aim 3, To further characterize regulatory mechanisms for stress responses mediated by the Mts1 bZIP protein: elucidate the role of the MAP kinase Spcl in controlling an autoinhibitory function of Mts1.

申请人的描述：ATF/CREB/AP-1家族的蛋白质是 监测细胞内和细胞外的信号转导途径 条件并将这些信号传输到下游目标。一些家庭 成员（例如 vFos 和 vJun）具有致癌性。 ATF/CREB/AP-1 蛋白共享一个 介导蛋白质二聚化和 序列特异性 DNA 结合活性。虽然 bZIP 域高度 bZIP 蛋白的一些组合彼此同源，形成二聚体， 而其他人则不然。此外，不同的bZIP二聚体具有显着的 区分密切相关的 DNA 结合位点的能力。这使得 一组有限的 bZIP 单体形成异二聚体库，每个 它可能调节涉及的一组特定基因的表达 在特定的生物反应中。了解 bZIP 蛋白 二聚体可以形成、它们占据哪些 DNA 位点以及如何调控此类事件 将揭示细胞稳态控制的基本方面和/或 差异化。裂殖酵母粟酒裂殖酵母提供了一个有吸引力的模型 进行此类研究的有机体。我们已经证明裂殖酵母的 bZIP 蛋白 在体内和体外形成组合二聚体。各种二聚体如下 MAP 激酶级联的控制和不同的二聚体引发不同的效应子 功能。因为粟酒裂殖酵母的基因组很小（大约 6,000 个基因），并且 测序项目已接近完成，可以识别并分析 大多数细胞 bZIP 蛋白。由于突变体缺乏 bZIP 蛋白并且 信号转导蛋白通常是可行的，也有可能 揭示上游调节和下游效应功能。一个 裂殖酵母bZIP蛋白二聚体的全面系统分析 被提议。目标 1，阐明粟酒裂殖酵母的 bZIP 字母表：确定 bZIP 二聚体的成对组合关联并鉴定 DNA 位点 它们以高亲和力结合。目标 2，揭示分子决定因素 组合二聚体形成和 DNA 结合：比较高分辨率 bZIP 二聚体单独的结构以及与其 DNA 位点复合的结构。目标 3，到 进一步表征由应激反应介导的调节机制 Mts1 bZIP 蛋白：阐明 MAP 激酶 Spcl 在控制 Mts1 的自抑制功能。