Light Signaling Connectivity in Arabidopsis

拟南芥中的光信号连接

• 批准号: 6831322
• 负责人: JOANNE CHORY
• 金额: $ 4.58万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6831322
• 关键词: Arabidopsis; RNA interference; South America; biological signal transduction; flavoproteins; functional /structural genomics; gene expression; gene mutation; genetic promoter element; genetic screening; microarray technology; molecular cloning; photobiology; plant genetics; plant growth /development; plant proteins; protein protein interaction

DESCRIPTION (provided by applicant): This research will be done primarily at the laboratory of Dr. Jorge J. Casal, University of Buenos Aires as an extension of NIH grant RO 1GM52413, Initial Events in Photoreceptor Signaling. The coordination of different signals controlling the transcriptome is essential for proper development of multicellular organisms. The long-term aim is to investigate the mechanisms of signaling connectivity by using light signaling in Arabidopsis thaliana as a model system. The system combines developmental complexity and sophisticated genetics with the ability to reversibly activate different receptors. By using Affymetrix full-genome chips we will identify transcripts whose abundance is controlled synergistically by the photoreceptors, phytochrome B (PHYB) and cryptochrome 1 (CRY1), in detailed kinetic experiments. A combination of unsupervised and supervised multivariate analysis of the data will be used to investigate the main patterns of the transcriptome. This information will provide a dynamic picture of the architecture of the network. The genes will be prioritized according to earliness and significance of the synergism and their function will be investigated by using reverse genetics, taking advantage of the publicly available large population of T-DNA mutants where disrupted genes have been identified. Complementary to this analysis, we will identify signaling elements necessary for the synergistic interaction between PHYB and CRY1 using a novel genetic screen. Seeds randomly mutagenized by T-DNA and populations mutagenized by chemical agents will be used for the screens. The mutated genes will be cloned either by sequencing the areas surrounding the T-DNA insert or by chromosome walking. Mutants resulting from forward or reverse genetics will be characterized by their ability to integrate signals when exposed to light combining different wavelengths. The mechanism by which different photoreceptors regulate an overlapping, yet diverse, set of developmental responses has been a long-standing question. Light signaling may use shared components. Alternatively, the signaling pathways may converge on a similar set of target genes. The experiments proposed here are designed as an integrated approach to begin answering this important question.

描述（由申请人提供）：这项研究将主要在布宜诺斯艾利斯大学Jorge J. Casal博士的实验室进行，作为NIH拨款RO 1GM52413，光感受器信号的初始事件的延伸。控制转录组的不同信号的协调对多细胞生物的正常发育至关重要。长期目标是利用拟南芥的光信号作为模型系统来研究信号连接的机制。该系统结合了发育复杂性和复杂的遗传学，以及可逆激活不同受体的能力。利用Affymetrix全基因组芯片，我们将在详细的动力学实验中鉴定出丰度受光感受器光敏色素B （PHYB）和隐色素1 （CRY1）协同控制的转录本。数据的无监督和有监督多变量分析的结合将用于研究转录组的主要模式。这些信息将提供网络体系结构的动态图像。这些基因将根据协同作用的早期性和重要性进行优先排序，它们的功能将通过使用反向遗传学进行研究，利用公开可用的大量T-DNA突变体，其中已鉴定出被破坏的基因。为了补充这一分析，我们将使用一种新的遗传筛选方法确定PHYB和CRY1之间协同相互作用所需的信号元件。T-DNA随机诱变的种子和化学药剂诱变的种群将用于筛选。突变基因将通过对T-DNA插入物周围区域进行测序或通过染色体行走来克隆。由正向或反向遗传产生的突变体的特点是，当暴露在不同波长的光下时，它们能够整合信号。不同的光感受器调控一套重叠而多样的发育反应的机制一直是一个长期存在的问题。光信令可以使用共享组件。或者，信号通路可能会聚在一组相似的靶基因上。这里提出的实验被设计成一个综合的方法来开始回答这个重要的问题。