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Centromeric Retroelements and Satellites Interact with Maize Kinetochore Protein CENH3

机译:着丝粒修饰元件和卫星与玉米动粒蛋白CENH3相互作用。

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摘要

Maize centromeres are composed of CentC tandem repeat arrays, centromeric retrotransposons (CRs), and a variety of other repeats. One particularly well-conserved CR element, CRM, occurs primarily as complete and uninterrupted elements and is interspersed thoroughly with CentC at the light microscopic level. To determine if these major centromeric DNAs are part of the functional centromere/kinetochore complex, we generated antiserum to maize centromeric histone H3 (CENH3). CENH3, a highly conserved protein that replaces histone H3 in centromeres, is thought to recruit many of the proteins required for chromosome movement. CENH3 is present throughout the cell cycle and colocalizes with the kinetochore protein CENPC in meiotic cells. Chromatin immunoprecipitation demonstrates that CentC and CRM interact specifically with CENH3, whereas knob repeats and Tekay retroelements do not. Approximately 38 and 33% of CentC and CRM are precipitated in the chromatin immunoprecipitation assay, consistent with data showing that much, but not all, of CENH3 colocalizes with CentC.
机译:玉米着丝粒由CentC串联重复序列阵列,着丝粒逆转录转座子(CRs)和各种其他重复序列组成。一种特别保存完好的CR元素CRM基本上是完整无间断的元素,并在光学显微镜下完全散布在CentC中。为了确定这些主要着丝粒DNA是否是功能着丝粒/线粒体复合体的一部分,我们生成了针对玉米着丝粒组蛋白H3(CENH3)的抗血清。 CENH3是一种高度保守的蛋白质,可替代着丝粒中的组蛋白H3,CENH3可以募集许多染色体运动所需的蛋白质。 CENH3存在于整个细胞周期中,并与减数分裂细胞中的线粒体蛋白CENPC共定位。染色质的免疫沉淀表明CentC和CRM与CENH3特异性相互作用,而球形重复和Tekay逆转录酶则没有。在染色质免疫沉淀试验中,约有38%和33%的CentC和CRM沉淀,与数据表明,有很多(但不是全部)的CENH3与CentC共定位的数据一致。

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