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首页> 外文期刊>Molecular biology of the cell >Identification of a novel 'chromosome scaffold' protein that associates with Tec elements undergoing En Masse elimination in Euplotes crassus
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Identification of a novel 'chromosome scaffold' protein that associates with Tec elements undergoing En Masse elimination in Euplotes crassus

机译：鉴定了一种新的“染色体支架”蛋白，该蛋白与Esplotes crassus中经历大规模消除的Tec元素相关

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摘要

During macronuclear development in the ciliate Euplotes crassus, the highly repetitive, transposon-like Tec elements possess an unusual chromatin structure. We observed that the Tec element chromatin is highly resistant to salt extraction and behaves like a nuclear matrix/chromosome scaffold-associated structure. Standard matrix/ scaffold extraction procedures identified two major proteins: 1) an similar to140-kDa protein that seems to be topoisomerase 11 based on its reactivity with anti-topoisomerase 11 antibodies, and 2) an 85-kDa protein that we further purified by acid extraction and have shown to be a novel protein by sequence analysis of its gene. The 85-kDa protein (p85) is a developmental stage-specific protein and is located exclusively in the developing macronucleus. Immunolocalization studies of p85 show that it colocalizes with topoisomerase II in chromatin. In addition, in situ hybridization combined with immunofluorescence localization of the proteins indicates that 100% of the Tec elements colocalize with 70% of the p85, whereas no significant colocalization with a total macronuclear sequence-specific probe is observed. p85 is the first developmental stage-specific protein identified as being specifically associated with sequences undergoing elimination in E. crassus. [References: 46]
机译：在纤毛Euplotes crassus的大核发育过程中，高度重复的转座子样Tec元素具有异常的染色质结构。我们观察到Tec元素染色质对盐提取具有高度抗性，并且表现得像核基质/染色体支架相关结构。标准基质/支架提取方法鉴定出两种主要蛋白质：1）一种类似于140-kDa的蛋白质，基于其与抗拓扑异构酶11抗体的反应性，似乎是拓扑异构酶11; 2）一种85-kDa的蛋白质，我们通过酸进一步纯化提取，并且通过对其基因的序列分析已显示是一种新型蛋白质。 85 kDa蛋白（p85）是一种发育阶段特异性蛋白，仅位于发育中的大核中。对p85的免疫定位研究表明，它与染色质中的拓扑异构酶II共定位。另外，蛋白质的原位杂交与免疫荧光定位相结合表明100％的Tec元件与70％的p85共同定位，而未观察到与总的大核序列特异性探针的明显共定位。 p85是第一个发育阶段特异性蛋白，被鉴定为与crassus crassus中经历消除的序列特异性结合。 [参考：46]

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