Structural and functional relationships of plant cryptochromes

植物隐花色素的结构和功能关系

• 批准号: 5420308
• 负责人: Professor Dr. Alfred Batschauer
• 金额: --
• 依托单位: AG Molekulare Pflanzenphysiologie und Photobiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2004
• 资助国家: 德国
• 起止时间: 2003-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5420308?language=en
• 关键词: Structural; functional; relationships; plant; cryptochromes

Cryptochromes are ubiquitous UV-A/blue-light photoreceptors related to light-driven DNA-repair enzymes (DNA-photolyases), which catalyze the removal of UV-B photoproducts from DNA. Cryptochromes, though having the same cofactors FAD and methenyl-tetrahydrofolate (MTHF) as the DNA-photolyases, have no photolyase activity. The crystal structures of three bacterial photolyases (Park et al., 1995; Tamada et al., 1997; Komori et al., 2001) and one bacterial cryptochrome have been solved (Brudler et al., 2003), demonstrating that this protein family is accessible to structural analysis at the atomic level. Most recently others and we discovered that cryptochromes can directly bind to DNA. Due to the progress we have made in the heterologous expression of plant cryptochromes we want to address in this proposal the function of plant cryptochromes at the atomic level. For this we will produce and analyze two cryptochromes that are located in different compartments in the plant cell, one located in the nucleus and the other located in the organelles. The proteins will be subjected to various spectroscopic analyses including crystallization and X-ray spectroscopy. Results from spectroscopy will be used to modify cryptochromes by site-directed mutagenesis and to analyze the effects of mutation on its biological function in vivo and in vitro.

隐色素是一种普遍存在的UV-A/蓝光感受器，与光驱动的DNA修复酶(DNA光解酶)有关，它催化从DNA中去除UV-B光产物。隐色素虽然具有与DNA光解酶相同的辅因子FAD和亚甲基四氢叶酸(MTHF)，但不具有光解酶活性。三个细菌光解酶(Park等人，1995；Tamada等人，1997；Komori等人，2001)和一个细菌隐色素(Brudler等人，2003)的晶体结构已经被解决，表明这个蛋白质家族可以在原子水平上进行结构分析。最近，其他人和我们发现，隐色素可以直接与DNA结合。由于我们在植物隐花色素的异源表达方面所取得的进展，我们希望在本提案中从原子水平上解决植物隐花色素的功能。为此，我们将生产和分析位于植物细胞不同隔室的两个隐色素，一个位于细胞核，另一个位于细胞器。这些蛋白质将接受各种光谱分析，包括结晶和X射线光谱。光谱学的结果将被用于通过定点突变来修饰隐花色素，并分析突变对其体内和体外生物学功能的影响。

项目成果

Recognition and repair of UV lesions in loop structures of duplex DNA by DASH-type cryptochrome

• DOI: 10.1073/pnas.0805830106
• 发表时间: 2008-12-30
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Pokorny, Richard;Klar, Tobias;Essen, Lars-Oliver
• 通讯作者: Essen, Lars-Oliver

Crystallization and preliminary X-ray analysis of cryptochrome 3 from Arabidopsis thaliana.

• DOI: 10.1107/s1744309105028897
• 发表时间: 2005-10
• 期刊: Acta crystallographica. Section F, Structural biology and crystallization communications
• 作者: R. Pokorný;T. Klar;L. Essen;A. Batschauer

Absorption and fluorescence spectroscopic characterization of cryptochrome 3 from Arabidopsis thaliana.

• DOI: 10.1016/j.jphotobiol.2006.03.007
• 发表时间: 2006-10
• 期刊: Journal of photochemistry and photobiology. B, Biology
• 作者: Sang-Hun Song;Bernhard Dick;A. Penzkofer;Richard Pokorny;A. Batschauer;L. Essen

Thermal degradation of (6R,S)-5,10-methenyltetrahydrofolate in aqueous solution at pH 8

(6R,S)-5,10-次甲基四氢叶酸在 pH 8 水溶液中的热降解

• DOI: 10.1016/j.chemphys.2009.01.005
• 发表时间: 2009
• 期刊: Chemical Physics
• 影响因子: 2.3
• 作者: Tyagi A;Penzkofer A;Batschauer A;Wolf E.
• 通讯作者: Wolf E.

Photocycle dynamics of the E149A mutant of cryptochrome 3 from Arabidopsis thaliana.

• DOI: 10.1016/j.jphotobiol.2009.08.005
• 发表时间: 2009-11
• 期刊: Journal of photochemistry and photobiology. B, Biology
• 作者: P. Zirak;A. Penzkofer;Julia Moldt;R. Pokorný;A. Batschauer;L. Essen