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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Cysteine modification of a specific repressor protein controls the translational status of nucleus-encoded LHCII mRNAs in Chlamydomonas
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Cysteine modification of a specific repressor protein controls the translational status of nucleus-encoded LHCII mRNAs in Chlamydomonas

机译：特定阻遏蛋白的半胱氨酸修饰控制衣原体中核编码的LHCII mRNA的翻译状态。

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摘要

The cytosolic RNA-binding protein NAB1 represses translation of LHCII (light-harvesting complex of photosystem II) encoding mRNAs by sequestration into translationally silent mRNP complexes in the green alga Chlamydomonas reinhardtii. NAB1 contains 2 cysteine residues, Cys-181 and Cys-226, within its C-terminal RRM motif. Modification of these cysteines either by oxidation or by alkylation in vitro was accompanied by a decrease in RNA-binding affinity for the target mRNA sequence. To confirm the relevance of reversible NAB1 cysteine oxidation for the regulation of its activity in vivo, we replaced both cysteines with serines. All examined cysteine single and double mutants exhibited a reduced antenna at PSII caused by a perturbed NAB1 deactivation mechanism, with double mutations and Cys-226 single mutations causing a stronger and more distinctive phenotype compared with the Cys-181 mutation. Our data indicated that the responsible redox control mechanism is mediated by modification of single cysteines. Polysome analyses and RNA co-immunoprecipitation experiments demonstrated the interconnection of the NAB1 thiol state and its activity as a translation repressor in vivo. NAB1 is fully active in its dithiol state and is reversibly deactivated by modification of its cysteines. In summary, this work is an example that cytosolic translation of nucleus encoded photosynthetic genes is regulated via a reversible cysteine-based redox switch in a RNA-binding translation repressor protein.
机译：胞质RNA结合蛋白NAB1通过在绿藻衣藻（Chlamydomonas reinhardtii）中螯合成翻译沉默的mRNP复合物，抑制了编码mRNA的LHCII（光系统II的光收集复合物）的翻译。 NAB1在其C端RRM基序内包含2个半胱氨酸残基Cys-181和Cys-226。这些半胱氨酸通过氧化或体外烷基化的修饰伴随着对靶mRNA序列的RNA结合亲和力的降低。为了确认可逆性NAB1半胱氨酸氧化与其体内活性调节的相关性，我们用丝氨酸替代了两个半胱氨酸。所有检测到的半胱氨酸单和双突变体均表现出受干扰的NAB1失活机制引起的PSII触角减小，与Cys-181突变相比，双突变和Cys-226单突变引起更强和更独特的表型。我们的数据表明负责任的氧化还原控制机制是由单个半胱氨酸的修饰介导的。多核糖体分析和RNA免疫共沉淀实验证明了NAB1硫醇状态的相互连接及其在体内作为翻译阻遏物的活性。 NAB1在其二硫醇状态下具有完全活性，并且通过其半胱氨酸的修饰可逆地失活。总而言之，这项工作是一个示例，该过程是通过RNA结合翻译阻遏蛋白中可逆的基于半胱氨​​酸的氧化还原开关调节核编码的光合基因的胞质翻译。

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    Lutz Wobbe; Olga Blifernez; Christian Schwarz; Jan H. Mussgnug; Joerg Nickelsen; Olaf Kruse;

  • 作者单位

    Department of Biology, University of Bielefeld, Algae BioTech Group, Universitaetsstrasse 25, D-33615 Bielefeld, Germany;

    Department of Biology, University of Bielefeld, Algae BioTech Group, Universitaetsstrasse 25, D-33615 Bielefeld, Germany;

    Ludwig-Maxmilians- Universitaet Muenchen, Department Biologie I-Molekulare Pflanzenwissenschaften, Grosshaderner Strasse 2-4, D-82152 Planegg-Martinsried, Germany;

    Department of Biology, University of Bielefeld, Algae BioTech Group, Universitaetsstrasse 25, D-33615 Bielefeld, Germany;

    Ludwig-Maxmilians- Universitaet Muenchen, Department Biologie I-Molekulare Pflanzenwissenschaften, Grosshaderner Strasse 2-4, D-82152 Planegg-Martinsried, Germany;

    Department of Biology, University of Bielefeld, Algae BioTech Group, Universitaetsstrasse 25, D-33615 Bielefeld, Germany;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    chlamydomonas reinhardtii; light harvesting antenna; redox control; translation control;

    机译：莱茵衣藻集光天线;氧化还原控制翻译控制;

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