A conserved lysine residue of plant Whirly proteins is necessary for higher order protein assembly and protection against DNA damage.
A conserved lysine residue of plant Whirly proteins is necessary for higher order protein assembly and protection against DNA damage.
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DOI：
10.1093/nar/gkr740
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发表时间：
2012-01
影响因子：
14.9
通讯作者：
Brisson N
中科院分区：
文献类型：
--
作者：
Cappadocia L;Parent JS;Zampini E;Lepage E;Sygusch J;Brisson N
所有生物都进化出了专门的DNA修复机制，以保护自己的基因组免受有害损伤，如DNA双链断裂。在植物细胞器中，这些损伤可以通过重组或通过微同源介导的断裂诱导复制途径来修复。在叶绿体和线粒体中，Whirly蛋白质都是这第二条途径的调节器。在这个精确的途径中，四聚体Whirly蛋白被认为与单链DNA结合，并防止切除的DNA分子与基因组中其他区域的虚假退火。在这项研究中，我们给这个模型增加了一层新的复杂性，通过原子力显微镜显示，马铃薯Whirly蛋白WHY2的四聚体在与长DNA分子结合时进一步组装成四聚体的六聚体，或24-MERS。这一过程依赖于K67介导的四聚体-四聚体相互作用，K67是植物Whirly蛋白中的一个高度保守的残基。该残基的突变取消了24-MERS的形成，而不影响蛋白质结构或与短DNA分子的结合。重要的是，我们表明，为这种赖氨酸突变的拟南芥Whirly蛋白无法挽救无Whirly突变植物对DNA双链断裂诱导剂的敏感性。
All organisms have evolved specialized DNA repair mechanisms in order to protect their genome against detrimental lesions such as DNA double-strand breaks. In plant organelles, these damages are repaired either through recombination or through a microhomology-mediated break-induced replication pathway. Whirly proteins are modulators of this second pathway in both chloroplasts and mitochondria. In this precise pathway, tetrameric Whirly proteins are believed to bind single-stranded DNA and prevent spurious annealing of resected DNA molecules with other regions in the genome. In this study, we add a new layer of complexity to this model by showing through atomic force microscopy that tetramers of the potato Whirly protein WHY2 further assemble into hexamers of tetramers, or 24-mers, upon binding long DNA molecules. This process depends on tetramer–tetramer interactions mediated by K67, a highly conserved residue among plant Whirly proteins. Mutation of this residue abolishes the formation of 24-mers without affecting the protein structure or the binding to short DNA molecules. Importantly, we show that an Arabidopsis Whirly protein mutated for this lysine is unable to rescue the sensitivity of a Whirly-less mutant plant to a DNA double-strand break inducing agent.
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影响因子：
11.8
作者：
Desveaux, D;Subramaniam, R;Brisson, N
通讯作者：
Brisson, N
DOI：
10.1073/pnas.89.17.8068
发表时间：
1992-09-01
影响因子：
11.1
作者：
CERUTTI, H;OSMAN, M;JAGENDORF, AT
通讯作者：
JAGENDORF, AT
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
3.6
作者：
Cosgriff S;Chintakayala K;Chim YT;Chen X;Allen S;Lovering AL;Grainger DC
通讯作者：
Grainger DC
影响因子：
7.2
作者：
Clough, SJ;Bent, AF
通讯作者：
Bent, AF