Functional analysis of the SYP1 syntaxin family

SYP1突触蛋白家族的功能分析

• 批准号: 5388416
• 负责人: Professor Dr. Gerd Jürgens
• 金额: --
• 依托单位: Abteilung für Entwicklungsgenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2002
• 资助国家: 德国
• 起止时间: 2001-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5388416?language=en
• 关键词: Functional; analysis; SYP1; syntaxin; family

Syntaxins are receptors on target membranes that mediate pathway-specific vesicle docking in intracellular trafficking. Among the 24 putative syntaxins of Arabidopsis, the 9 members of the SYP1 family display weak similarity to plasma membrane-associated syntaxins of other eukaryotes. Except for the cytokinesis-specific syntaxin KNOLLE (SYP111) and a putative orthologue of the plasma membrane-localised tobacco SYR1 (SYP121), nothing is known about SYP1 proteins. The aim of this project is to analyse all members of the SYP1 family. We will search for knockout mutants and examine their possible defects in intracellular trafficking. Expression and subcellular localisation of myc-tagged SYP1 proteins will be studied in transgenic plants. Myc-tagged SYP1 proteins will be expressed under the control of KNOLLE (and SYR1) cis-regulatory sequences to assess their functional specificity in terms of targeting to the cell plate (and plasma membrane) and ability to cross-complement knolle (and syr1) mutants. Using the cell cycle-regulated accumulation, targeting and function of KNOLLE protein as an assay system, we aim to identify functional domains of SYP1 proteins. To this end, chimaeric proteins consisting of varying proportions of KNOLLE and other SYP1 proteins will be analysed for targeting, stability and knolle rescuing activity.

突触融合蛋白是靶膜上的受体，介导细胞内运输中的路径特异性囊泡对接。在拟南芥的24个推定的突触融合蛋白中，SYP1家族的9个成员与其他真核生物的质膜相关突触融合蛋白显示出微弱的相似性。除了烟草分裂特异性突触融合蛋白诺勒（SYP111）和质膜定位的烟草SYR1（SYP121）的推定直向同源物外，对SYP1蛋白一无所知。该项目的目的是分析SYP1家族的所有成员。我们将寻找敲除突变体，并检查它们在细胞内运输中可能存在的缺陷。将在转基因植物中研究myc标记的SYP1蛋白的表达和亚细胞定位。Myc标记的SYP1蛋白将在诺勒（和SYR1）顺式调节序列的控制下表达，以评估其靶向细胞板（和质膜）的功能特异性和交叉互补诺勒（和SYR1）突变体的能力。利用细胞周期调控的诺勒蛋白的积累、靶向和功能作为检测系统，我们的目的是鉴定SYP1蛋白的功能结构域。为此，将分析由不同比例的诺勒和其他SYP1蛋白组成的嵌合蛋白的靶向、稳定性和小诺勒拯救活性。