Protein components of mitochondrial DNA nucleoids in higher eukaryotes
Protein components of mitochondrial DNA nucleoids in higher eukaryotes
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DOI：
10.1074/mcp.m300035-mcp200
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发表时间：
2003-11-01
影响因子：
7
通讯作者：
Kobayashi, R
中科院分区：
文献类型：
--
作者：
Bogenhagen, DF;Wang, YS;Kobayashi, R
线粒体 DNA (mtDNA) 不包装在核小体颗粒中，但据报道与线粒体内膜结合。用非离子去污剂温和裂解非洲爪蟾卵母细胞线粒体，释放出一种核蛋白复合物，其中含有与先前表征的 DNA 结合伴侣、线粒体转录因子 A (mtTFA) 以及测序鉴定的一系列内膜蛋白相关的 mtDNA。更广泛的去污剂处理从DNA中剥离了大部分这些蛋白质，在类核核心中留下了有限数量的蛋白质。对与 mtDNA 相关的主要蛋白质进行测序，揭示了预期的 mtDNA 结合蛋白、mtTFA 和线粒体单链 DNA 结合蛋白 (mtSSB)，以及之前未报道的与 mtDNA 相关的四种蛋白质。这些包括腺嘌呤核苷酸转位子 1、丙酮酸脱氢酶的含硫辛酰的 E2 亚基和支链 α-酮酸脱氢酶和抑制素 2。这些蛋白质中的几种与含 mtTFA 的 mtDNA 核仁的关联通过免疫沉淀得到证实。
Mitochondrial DNA ( mtDNA) is not packaged in nucleosomal particles, but has been reported to associate with the mitochondrial inner membrane. Gentle lysis of Xenopus oocyte mitochondria with nonionic detergent liberates a nucleoprotein complex containing mtDNA associated with a previously characterized DNA binding partner, mitochondrial transcription factor A ( mtTFA), as well as a series of inner membrane proteins identified by sequencing. More extensive detergent treatment stripped most of these proteins from the DNA, leaving a limited number of proteins in a nucleoid core. Sequencing of the major proteins retained in association with mtDNA revealed the expected mtDNA binding proteins, mtTFA and mitochondrial single-stranded DNA binding protein ( mtSSB), as well as four proteins not previously reported to associate with mtDNA. These include adenine nucleotide translocator 1, the lipoyl-containing E2 subunits of pyruvate dehydrogenase and branched chain alpha-ketoacid dehydrogenase and prohibitin 2. The association of several of these proteins with mtTFA-containing mtDNA nucleoids was confirmed by immunoprecipitation.