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A Conserved Mg2+-Dependent Exonuclease Degrades Organelle DNA during Arabidopsis Pollen Development

机译:在拟南芥花粉发育过程中一个保守的Mg2 +依赖性核酸外切酶会降解细胞器DNA。

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摘要

In plant cells, mitochondria and plastids contain their own genomes derived from the ancestral bacteria endosymbiont. Despite their limited genetic capacity, these multicopy organelle genomes account for a substantial fraction of total cellular DNA, raising the question of whether organelle DNA quantity is controlled spatially or temporally. In this study, we genetically dissected the organelle DNA decrease in pollen, a phenomenon that appears to be common in most angiosperm species. By staining mature pollen grains with fluorescent DNA dye, we screened Arabidopsis thaliana for mutants in which extrachromosomal DNAs had accumulated. Such a recessive mutant, termed defective in pollen organelle DNA degradation1 (dpd1), showing elevated levels of DNAs in both plastids and mitochondria, was isolated and characterized. DPD1 encodes a protein belonging to the exonuclease family, whose homologs appear to be found in angiosperms. Indeed, DPD1 has Mg2+-dependent exonuclease activity when expressed as a fusion protein and when assayed in vitro and is highly active in developing pollen. Consistent with the dpd phenotype, DPD1 is dual-targeted to plastids and mitochondria. Therefore, we provide evidence of active organelle DNA degradation in the angiosperm male gametophyte, primarily independent of maternal inheritance; the biological function of organellar DNA degradation in pollen is currently unclear.
机译:在植物细胞中,线粒体和质体含有其自身的基因,这些基因组来自祖先细菌内共生体。尽管它们的遗传能力有限,这些多拷贝细胞器基因组仍占细胞总DNA的很大一部分,这引起了细胞器DNA数量是在空间上还是时间上受到控制的问题。在这项研究中,我们通过遗传解剖了花粉中细胞器DNA的减少,这种现象在大多数被子植物物种中似乎很常见。通过用荧光DNA染料对成熟的花粉粒染色,我们筛选了拟南芥中积累了染色体外DNA的突变体。分离并鉴定了一种隐性突变体,该突变体在花粉细胞器DNA降解1(dpd1)中表现出缺陷,显示质体和线粒体中的DNA含量均升高。 DPD1编码一种属于核酸外切酶家族的蛋白质,其同源物似乎存在于被子植物中。的确,当DPD1表达为融合蛋白并在体外测定时,它具有Mg 2 + 依赖的核酸外切酶活性,并且在花粉发育中具有很高的活性。与dpd表型一致,DPD1双重靶向质体和线粒体。因此,我们提供了被子植物雄配子体中活性细胞器DNA降解的证据,主要独立于母体遗传。目前尚不清楚花粉中的细胞器DNA降解的生物学功能。

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