Molecular Analysis of the rbcS Multigene Family

rbcS 多基因家族的分子分析

• 批准号: 8905282
• 负责人: Wilhelm Gruissem
• 金额: $ 32.78万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8905282&HistoricalAwards=false
• 关键词: Molecular; Analysis; rbcS; Multigene; Family

This project will investigate the molecular mechanism involved in the differential expression of the multigene family members for the small subunit (SSU) of ribulose.1, 5.bisphosphate carboxylase (rbcS genes) using tomato as a model system. Tomato has five rbcs genes rbcS.1, rbcS.2, rbcS.3A and rbcS.3C, which encode three iso.variant SSU proteins. The five rbcS genes are differentially transcribed in cotyledons, leaves and fruit. Several 5' DNA sequences which are functionally undefined are conserved among the five tomato rbcS genes which share similar developmental and organ.specific expression pattern. In addition, mRNAs from the individual tomato rbcS genes accumulate to substantially different levels in different tissues, but this pattern is not reflected in the accumulation of the three iso. variant SSU proteins. We will examine rbcS cis.acting DNA regions to define their role in the developmental and organ. specific on.off control of transcription and examine how they interact. We will identify protein.binding domains by DNasel footprint analysis to determine if the differential transcription of the rbcS genes is correlated with differences in the DNA: protein binding pattern of 5' DNA fragments. This will complement our analysis of cis.acting DNA sequences in transgenic plants and transient assays. Furthermore, transcription run.on assays and quantitative hybridizations will be used to assess the extent to which transcriptional regulation of the five rbcS genes establishes the relative levels of their mRNAs. At the translational level, we will test if variations in the length of the rbcS 5' untranslated regions result in the differential accumulation of iso.variant SSU proteins. To attain further information on the role of differential rbcS express in fruit, we will take advantage of a fruit.specific promoter for the production of antisense.RNA to block rbcS.1 and rbcS.2 expression. Finally, we will activate the normally inactive rbcS.3A gene to produce the iso.variant 3 SSU protein in fruit to determine what effect, if any, this has on organ development. Developmental, organ.specific and light.dependent regulation of transcription and translation are important control steps in the expression of nuclear genes encoding photosynthetic proteins found in the chloroplasts of higher plants. The results of this study should provide greater insights into those processes.

本项目将研究参与的分子机制， 多基因家族成员的差异表达， 核酮糖1，5-二磷酸羧化酶小亚基（SSU (rbcS基因）使用番茄作为模型系统。 番茄有五个 rbcs基因rbcS.1、rbcS.2、rbcS.3A和rbcS.3C，其编码 三种异源变异SSU蛋白。 这五个rbcS基因是 在子叶、叶和果实中差异转录。 几个功能上不确定的5' DNA序列是 在5个番茄rbcS基因中保守， 发育和器官特异性表达模式。 在 此外，来自单个番茄rbcS基因的mRNA积累 在不同的组织中有显著不同的水平，但是， 模式是不反映在积累的三个iso。 变异SSU蛋白。 我们将检测rbcS顺式作用DNA， 各区域确定其在发展和机构中的作用。 具体的开关控制转录，并检查他们如何 互动. 我们将通过DNaseI鉴定蛋白质结合结构域 足迹分析以确定差异转录 rbcS基因的差异与DNA的差异有关： 5 ′ DNA片段的蛋白结合模式。 这将 补充了我们对转基因植物中顺式作用DNA序列的分析， 植物和瞬时测定。 此外，转录运行。 检测和定量杂交将用于评估 五个rbcS基因的转录调控程度 确定它们的mRNA的相对水平。 在 翻译水平，我们将测试，如果在长度的变化， rbcS 5'端非翻译区导致了 积累同种变体SSU蛋白。 为了进一步实现 关于差异rbcS在果实中表达的作用的信息，我们 将利用水果特异性启动子， 产生反义RNA以阻断rbcS.1和rbcS.2 表情 最后，我们将激活通常不活动的 rbcS.3A基因在果实中产生同种变体3 SSU蛋白， 确定这对器官发育有什么影响，如果有的话。 发育的、器官的、特定的和光的依赖性调节 转录和翻译是转录过程中重要的控制步骤。 光合蛋白核基因的表达 存在于高等植物的叶绿体中。 的结果 研究应使人们更深入地了解这些进程。