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首页> 外文期刊>Journal of anti-aging medicine >A telomere-binding protein (TRF2/MTBP) from mouse nuclear matrix with motives of an intermediate filament-type rod domain.
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A telomere-binding protein (TRF2/MTBP) from mouse nuclear matrix with motives of an intermediate filament-type rod domain.

机译：来自小鼠核基质的端粒结合蛋白（TRF2 / MTBP），具有中间丝状杆状结构域的动机。

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摘要

In previous work, we identified a telomeric DNA-binding protein (termed telomere-membrane binding protein, MTBP) in the envelope of the frog oocyte nucleus and raised antibodies against it. Here we present immunological evidence which suggests strongly that MTBP is identical with the vertebrate telomeric DNA-binding protein TRF2 (telomere-repeat factor 2). MTBP/TRF2 possesses motif which resembles rod domain characteristic of intermediate filament (IF) proteins as shown by immunological cross-reactivity with characteristic antibodies, as well as amino acid sequence homology. Anti-MTBP antibodies recognised a protein of the same M, as TRF2 in extracts of mouse nuclei and nuclear matrix as shown by ion-exchange chromatography, gel shift assays, and Western blots. This mouse MTBP analogue forms more stable complexes with the vertebrate telomeric DNA fragment (T(2)AG(3))(135) than with the corresponding fragment from Tetrahymena (T(2)G(4))(130). Proteins in each of these complexes are recognised by anti-MTBP antibody. In situ hybridization with the vertebrate telomeric DNA sequence (T(2)AG(3))(135) and immunofluorescence with anti-MTBP antibody had shown earlier that these are co-localised in the nucleus of mouse cells, and here MTBP is shown to be associated with the residual membrane of hepatocyte nuclei using Western blotting and immunofluorescence. Some immunofluorescence signal from MTBP is localized at chromosome extremities on metaphase plates from mouse cell culture, but the main signal is seen in patches scattered around the chromosomes which were identified as remnants of the nuclear envelope by double labelling with antibodies against lamin B. These observations suggest that MTBP/TRF2 is a good candidate for the attachment of telomeres to the nuclear envelope in somatic cells.
机译：在以前的工作中，我们在青蛙卵母细胞核的被膜中鉴定了端粒DNA结合蛋白（称为端粒膜结合蛋白，MTBP），并提出了针对它的抗体。在这里，我们提供了免疫学证据，这些证据强烈表明MTBP与脊椎动物端粒DNA结合蛋白TRF2（端粒重复因子2）相同。 MTBP / TRF2具有类似于中间丝（IF）蛋白杆结构域特征的基序，如与特征性抗体的免疫交叉反应以及氨基酸序列同源性所示。抗MTBP抗体在小鼠细胞核和核基质提取物中识别出与TRF2相同M的蛋白质，如离子交换色谱法，凝胶迁移测定法和Western印迹法所示。该小鼠MTBP类似物与脊椎动物端粒DNA片段（T（2）AG（3））（135）形成的稳定复合物比与四膜虫（T（2）G（4））（130）的相应片段形成的稳定复合物更复杂。这些复合物中的每一种中的蛋白质均被抗MTBP抗体识别。与脊椎动物的端粒DNA序列（T（2）AG（3））（135）进行原位杂交并用抗MTBP抗体进行免疫荧光检测表明，这些蛋白共定位于小鼠细胞核中，此处显示MTBP通过Western印迹和免疫荧光分析发现其与肝细胞核的残留膜相关。来自MTBP的一些免疫荧光信号位于小鼠细胞培养中期板的染色体末端，但是主要信号出现在散布在染色体周围的斑块中，这些斑块通过用抗lamin B抗体进行双重标记而被识别为核被膜的残留物。提示MTBP / TRF2是端粒附着于体细胞核膜的良好候选者。

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