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Sll0606 蛋白是集胞藻 PCC 6803 中光系统 II 组装/稳定性所必需的。

The Sll0606 protein is required for photosystem II assembly/stability in the cyanobacterium Synechocystis sp. PCC 6803.

机构信息

Division of Biochemistry and Molecular Biology, Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana 70803, USA.

出版信息

J Biol Chem. 2010 Oct 15;285(42):32047-54. doi: 10.1074/jbc.M110.166983. Epub 2010 Aug 19.

Abstract

An insertional transposon mutation in the sll0606 gene was found to lead to a loss of photoautotrophy but not photoheterotrophy in the cyanobacterium Synechocystis sp. PCC 6803. Complementation analysis of this mutant (Tsll0606) indicated that an intact sll0606 gene could fully restore photoautotrophic growth. Gene organization in the vicinity of sll0606 indicates that it is not contained in an operon. No electron transport activity was detected in Tsll0606 using water as an electron donor and 2,6-dichlorobenzoquinone as an electron acceptor, indicating that Photosystem II (PS II) was defective. Electron transport activity using dichlorophenol indolephenol plus ascorbate as an electron donor to methyl viologen, however, was the same as observed in the control strain. This indicated that electron flow through Photosystem I was normal. Fluorescence induction and decay parameters verified that Photosystem II was highly compromised. The quantum yield for energy trapping by Photosystem II (F(V)/F(M)) in the mutant was less than 10% of that observed in the control strain. The small variable fluorescence yield observed after a single saturating flash exhibited aberrant Q(A)(-) reoxidation kinetics that were insensitive to dichloromethylurea. Immunological analysis indicated that whereas the D2 and CP47 proteins were modestly affected, the D1 and CP43 components were dramatically reduced. Analysis of two-dimensional blue native/lithium dodecyl sulfate-polyacrylamide gels indicated that no intact PS II monomer or dimers were observed in the mutant. The CP43-less PS II monomer did accumulate to detectable levels. Our results indicate that the Sll0606 protein is required for the assembly/stability of a functionally competent Photosystem II.

摘要

在集胞藻 PCC 6803 中,发现 sll0606 基因的插入转座子突变导致其丧失了光自养能力,但没有丧失光异养能力。对该突变体(Tsll0606)的互补分析表明,一个完整的 sll0606 基因可以完全恢复光自养生长。sll0606 附近的基因组织表明它不包含在一个操纵子中。在 Tsll0606 中,使用水作为电子供体和 2,6-二氯苯醌作为电子受体时,没有检测到电子传递活性,表明 PSII(光合系统 II)有缺陷。然而,使用二氯苯酚吲哚酚加抗坏血酸作为电子供体到甲基紫精时,电子传递活性与对照菌株相同。这表明电子流经 PSI 是正常的。荧光诱导和衰减参数验证 PSII 高度受损。突变体中 PSII 的能量捕获量子产率(F(V)/F(M))小于对照菌株观察到的 10%。在单个饱和闪光后观察到的小可变荧光产率表现出异常的 Q(A)(-)再氧化动力学,对二氯甲基脲不敏感。免疫分析表明,虽然 D2 和 CP47 蛋白受到轻微影响,但 D1 和 CP43 组件显著减少。二维蓝色 native/十二烷基硫酸钠-聚丙烯酰胺凝胶分析表明,在突变体中没有观察到完整的 PSII 单体或二聚体。CP43 缺失的 PSII 单体确实积累到可检测水平。我们的结果表明,Sll0606 蛋白是 PSII 组装/稳定性所必需的。

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