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Chromodomain protein Tcd1 is required for macronuclear genome rearrangement and repair in Tetrahymena

机译:四面膜大分子基因组重排和修复需要染色体结构域蛋白Tcd1

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摘要

The survival of an organism’s progeny depends on the maintenance of its genome. Programmed DNA rearrangement and repair in Tetrahymena occur during the differentiation of the developing somatic macronuclear genome from the germ line micronuclear genome. Tetrahymena chromodomain protein (Tcd1) exhibited dynamic localization from the parental to the developing macronuclei. In the developing macronuclei, Tcd1 colocalized with Pdd1 and H3K9me3. Furthermore, Tcd1 colocalized with Pdd1 in the conjusome and “donut structure” of DNA elimination heterochromatin region. During the growth and conjugation stages, TCD1 knockout cells appeared normal and similar to wild-type strains. In addition, these knockout cells proceeded to the 2MAC-1MIC stage. However, the progeny of the TCD1 knockout cells did not grow upon return to SPP medium and eventually died. The deletion of the internal elimination sequence R element was partially disrupted in the developing new macronuclei. Gamma H2A staining showed that Tcd1 loss induced the accumulation of DNA double-strand breaks and the failure of genome repair. These results suggest that the chromodomain protein Tcd1 is required for the rearrangement and repair of new macronuclear genome in Tetrahymena.
机译:生物体后代的生存取决于其基因组的维持。四膜虫的程序化DNA重排和修复发生在发育中的体细胞大核基因组与种系微核基因组的分化过程中。四膜虫色域蛋白(Tcd1)表现出从亲代到发育中的大核的动态定位。在发育中的大核中,Tcd1与Pdd1和H3K9me3共定位。此外,Tcd1与Pdd1共定位于DNA消除异染色质区域的缀合物和“甜甜圈结构”中。在生长和结合阶段,TCD1敲除细胞似乎正常且与野生型菌株相似。此外,这些敲除细胞进入了2MAC-1MIC阶段。但是,TCD1基因敲除细胞的后代在返回SPP培养基后并未生长,最终死亡。内部消除序列R元件的缺失在正在发育的新大核中被部分破坏。伽马H2A染色显示Tcd1丢失导致DNA双链断裂的积累和基因组修复的失败。这些结果表明,色域蛋白Tcd1是四膜虫中新的大核基因组的重排和修复所必需的。

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