Rurek Michal, Woyda-Ploszczyca Andrzej M, Jarmuszkiewicz Wieslawa
Department of Cellular and Molecular Biology, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University in Poznań, Umultowska 89, 61-614 Poznań, Poland.
Department of Bioenergetics, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University in Poznań, Umultowska 89, 61-614 Poznań, Poland.
Biochim Biophys Acta. 2015 Apr-May;1847(4-5):399-417. doi: 10.1016/j.bbabio.2015.01.005. Epub 2015 Jan 21.
The biogenesis of the cauliflower curd mitochondrial proteome was investigated under cold, heat and the recovery. For the first time, two dimensional fluorescence difference gel electrophoresis was used to study the plant mitochondrial complexome in heat and heat recovery. Particularly, changes in the complex I and complex III subunits and import proteins, and the partial disintegration of matrix complexes were observed. The presence of unassembled subunits of ATP synthase was accompanied by impairment in mitochondrial translation of its subunit. In cold and heat, the transcription profiles of mitochondrial genes were uncorrelated. The in-gel activities of respiratory complexes were particularly affected after stress recovery. Despite a general stability of respiratory chain complexes in heat, functional studies showed that their activity and the ATP synthesis yield were affected. Contrary to cold stress, heat stress resulted in a reduced efficiency of oxidative phosphorylation likely due to changes in alternative oxidase (AOX) activity. Stress and stress recovery differently modulated the protein level and activity of AOX. Heat stress induced an increase in AOX activity and protein level, and AOX1a and AOX1d transcript level, while heat recovery reversed the AOX protein and activity changes. Conversely, cold stress led to a decrease in AOX activity (and protein level), which was reversed after cold recovery. Thus, cauliflower AOX is only induced by heat stress. In heat, contrary to the AOX activity, the activity of rotenone-insensitive internal NADH dehydrogenase was diminished. The relevance of various steps of plant mitochondrial biogenesis to temperature stress response and recovery is discussed.
研究了花椰菜球状体线粒体蛋白质组在冷、热及恢复条件下的生物发生过程。首次利用二维荧光差异凝胶电泳研究了热胁迫及热恢复过程中植物线粒体复合物组。特别地,观察到复合物I和复合物III亚基及导入蛋白的变化,以及基质复合物的部分解体。ATP合酶未组装亚基的存在伴随着其亚基线粒体翻译的受损。在冷胁迫和热胁迫下,线粒体基因的转录谱不相关。应激恢复后,呼吸复合物的凝胶内活性受到特别影响。尽管热胁迫下呼吸链复合物总体稳定,但功能研究表明其活性和ATP合成产量受到影响。与冷胁迫相反,热胁迫导致氧化磷酸化效率降低,可能是由于交替氧化酶(AOX)活性的变化。胁迫和胁迫恢复对AOX的蛋白水平和活性有不同的调节作用。热胁迫诱导AOX活性、蛋白水平以及AOX1a和AOX1d转录水平增加,而热恢复则逆转了AOX蛋白和活性的变化。相反,冷胁迫导致AOX活性(和蛋白水平)降低,冷恢复后则逆转。因此,花椰菜AOX仅由热胁迫诱导。在热胁迫下,与AOX活性相反,鱼藤酮不敏感的内部NADH脱氢酶的活性降低。讨论了植物线粒体生物发生的各个步骤与温度胁迫响应及恢复的相关性。