Light harvesting proteins in diatoms - functional heterogeneity and regulation

硅藻中的光捕获蛋白 - 功能异质性和调节

• 批准号: 146946584
• 负责人: Professorin Dr. Claudia Büchel
• 金额: --
• 依托单位: Institut für Molekulare Biowissenschaften
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/146946584?language=en
• 关键词: Light; harvesting; proteins; diatoms; functional

The major aim of the project is to examine more deeply the effects of the newly identified, unusual cryptochrome CryP in the pennate diatom Phaeodactylum tricornutum. CryP, having DASH like features according to spectroscopic analyses but also grouping close to plant Crys in sequence analyses, was shown to be involved in the regulation of the overall protein levels of different light-harvesting proteins (fucoxanthin chlorophyll proteins, FCPs). Here, we will focus on the influence of CryP on the organisation of light harvesting proteins. Many FCPs are expressed in diatoms and assemble mostly in trimers, which differ in polypeptide composition and also slightly in pigmentation. We want to use our CryP knockdown lines to test biochemically, which FCP trimers are influenced by the action of CryP. Whether this influence is in relation to the circadian clock will be determined on mRNA level, since CryP is related to plant Crys. In addition, the cellular localization of CryP should be clarified. Since the spectroscopic features of CryP resemble CRY-DASH proteins, its possible bi-functionality, i.e. an involvement in DNA repair, will be tested as well. Spectroscopic and structural investigations on the protein together with mRNA quantifications under different light conditions should provide further insight into the mode of action, as well as the identification of possible interaction partners.

该项目的主要目的是更深入地研究新发现的、不寻常的隐色素CRYP在羽状硅藻三角褐指藻中的作用。根据光谱分析，CRYP具有短划线的特征，但在序列分析中也与植物Crys相似，它被证明参与了不同捕光蛋白(岩藻黄质-叶绿素蛋白，FCP)的整体蛋白质水平的调节。在这里，我们将集中在CRYP对光捕获蛋白组织的影响。许多FCP在硅藻中表达，并主要以三聚体形式组装，三聚体的多肽组成略有不同，色素也略有不同。我们希望使用我们的CRYP基因敲除系来进行生化测试，FCP三聚体受CRYP的作用影响。这种影响是否与生物钟有关将在mRNA水平上确定，因为CRYP与植物Crys有关。此外，CRYP的细胞定位还需进一步阐明。由于CRYP的光谱特征类似于Crydash蛋白，它可能的双功能，即参与DNA修复，也将被测试。对蛋白质的光谱和结构研究，以及不同光条件下的mRNA定量，应该可以进一步深入了解作用模式，以及识别可能的相互作用伙伴。