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首页> 外文期刊>Molecular and Cellular Biology >Nucleus-encoded histone H1-like proteins are associated with kinetoplast DNA in the trypanosomatid Crithidia fasciculata.
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Nucleus-encoded histone H1-like proteins are associated with kinetoplast DNA in the trypanosomatid Crithidia fasciculata.

机译：核编码的组蛋白H1样蛋白与锥虫锥虫Crithidia fasciculata中的运动塑料DNA相关。

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Kinetoplast DNA (kDNA), the mitochondrial DNA of trypanosomatids, consists of thousands of minicircles and 20 to 30 maxicircles catenated into a single large network and exists in the cell as a highly organized compact disc structure. To investigate the role of kinetoplast-associated proteins in organizing and condensing kDNA networks into this disc structure, we have cloned three genes encoding kinetoplast-associated proteins. The KAP2, KAP3, and KAP4 genes encode proteins p18, p17, and p16, respectively. These proteins are small basic proteins rich in lysine and alanine residues and contain 9-amino-acid cleavable presequences. Proteins p17 and p18 are closely related to each other, with 48% identical residues and carboxyl tails containing almost exclusively lysine, alanine, and serine or threonine residues. These proteins have been expressed as Met-His6-tagged recombinant proteins and purified by metal chelate chromatography. Each of the recombinant proteins is capable of compacting kDNA networks in vitro and was shown to bind preferentially to a specific fragment of minicircle DNA. Expression of each of these proteins in an Escherichia coli mutant lacking the HU protein rescued a defect in chromosome condensation and segregation in the mutant cells and restored a near-normal morphological appearance. Proteins p16, p17, and p18 have been localized within the cell by immunofluorescence methods and appear to be present throughout the kDNA. Electron-microscopic immunolocalization of p16 shows that p16 is present both within the kDNA disc and in the mitochondrial matrix at opposite edges of the kDNA disc. Our results suggest that nucleus-encoded H1-like proteins may be involved in the organization and segregation of kDNA networks in trypanosomatids.
机译：锥虫线粒体的线粒体DNA运动塑料DNA（kDNA）由数以千计的小圆和20至30个最大圆组成，它们连接成一个大型网络，并以高度组织的光盘结构存在于细胞中。为了研究动塑料相关蛋白在组织和浓缩kDNA网络到该盘结构中的作用，我们克隆了三个编码动塑料相关蛋白的基因。 KAP2，KAP3和KAP4基因分别编码蛋白p18，p17和p16。这些蛋白质是富含赖氨酸和丙氨酸残基的小碱性蛋白质，并含有9个氨基酸可切割的前序列。蛋白质p17和p18彼此密切相关，具有48％相同的残基和羧基尾部，几乎仅包含赖氨酸，丙氨酸和丝氨酸或苏氨酸残基。这些蛋白已表达为带有Met-His6标签的重组蛋白，并通过金属螯合色谱法纯化。每个重组蛋白都能够在体外压缩kDNA网络，并显示出优先结合微环DNA的特定片段的能力。这些蛋白在缺少HU蛋白的大肠杆菌突变体中的表达挽救了突变体细胞染色体浓缩和分离的缺陷，并恢复了接近正常的形态外观。蛋白p16，p17和p18已通过免疫荧光方法定位在细胞内，并且似乎存在于整个kDNA中。 p16的电子显微镜免疫定位显示，p16既存在于kDNA圆盘内，也存在于kDNA圆盘相对边缘的线粒体基质中。我们的结果表明，核编码的H1样蛋白可能参与了锥虫的kDNA网络的组织和分离。

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