Kinetoplast maxicircle DNA replication in Crithidia fasciculata and Trypanosoma brucei

Kinetoplast maxicircle DNA replication in Crithidia fasciculata and Trypanosoma brucei
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Crithidia fasciculata 和 Trypanosoma brucei 中动质体上环 DNA 复制

DOI:
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发表时间:
1995
影响因子:
5.3
通讯作者:
P. T. Englund
P. T. Englund
中科院分区:
生物学2区
文献类型:
--
作者:
Laura Rocco Carpenter;P. T. Englund

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着丝体DNA,即锥虫的线粒体DNA,由几千个小环和几十个大环组成,它们在拓扑结构上互锁在一个巨大的网络中。我们研究了束状棘虫和布鲁氏锥虫的大环DNA复制,用电子显微镜分析了它们的复制中间体。通过在体内用4,5',8-三甲基补骨脂素和紫外线辐射引入DNA链间交联来稳定复制中间体的分支迁移。由拓扑异构酶II对着丝体DNA网络进行十烷化而产生的单个最大环的电子显微镜显示完整的最大环θ结构。用限制性内切酶切割线性化的最大环DNA的分析显示,分支复制中间体来源于θ结构。对这两种寄生虫的线性化分支分子的测量表明,复制始于可变区(以重复序列为特征的非编码片段),并在标准图谱上沿顺时针方向单向进行。
Kinetoplast DNA, the mitochondrial DNA of trypanosomatids, is composed of several thousand minicircles and a few dozen maxicircles, all of which are topologically interlocked in a giant network. We have studied the replication of maxicircle DNA, using electron microscopy to analyze replication intermediates from both Crithidia fasciculata and Trypanosoma brucei. Replication intermediates were stabilized against branch migration by introducing DNA interstrand cross-links in vivo with 4,5',8-trimethylpsoralen and UV radiation. Electron microscopy of individual maxicircles resulting from a topoisomerase II decatenation of kinetoplast DNA networks revealed intact maxicircle theta structures. Analysis of maxicircle DNA linearized by restriction enzyme cleavage revealed branched replication intermediates derived from theta structures. Measurements of the linearized branched molecules in both parasites indicate that replication initiates in the variable region (a noncoding segment characterized by repetitive sequences) and proceeds unidirectionally, clockwise on the standard map.
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发表时间: 1987-05
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影响因子: --
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发表时间: 1992
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