Biochemical characterization and comparison of two closely related active mariner transposases.
Biochemical characterization and comparison of two closely related active mariner transposases.
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DOI：
10.1021/bi401193w
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发表时间：
2014-02-04
期刊：
影响因子：
2.9
通讯作者：
Richardson, Julia M.
中科院分区：
文献类型：
--
作者：
Trubitsyna, Maryia;Morris, Elizabeth R.;Finnegan, David J.;Richardson, Julia M.
大多数DNA转座子通过剪切和粘贴机制从一个基因组位置移动到另一个基因组位置，并且是基因组操作的有用工具。标记转座子每个末端的短反向重复（IR）DNA序列被DNA转座酶（由转座子本身编码）识别。这种酶切割转座子末端并将它们整合到新的基因组位置。我们在这里报告的生物物理和生化特性的比较密切相关的和活跃的水手/Tc 1家族DNA转座酶：Mboumar-9和Mos 1。我们比较了酶对它们自己的IR序列的体外切割活性，以及它们的反向重复序列的交叉识别。我们发现，像Mos 1，未标记的重组Mboumar-9转座酶是一个二聚体，并形成一个稳定的复合物与反向重复DNA的存在下，镁离子。Mboumar-9转座酶以Mos 1转座酶观察到的方式切割其反向重复DNA。Mos 1和Mboumar-9转座酶之间IR序列的交叉识别最小，尽管这些酶具有68%相同的氨基酸序列。转座酶共享共同的生物物理和生物化学特性，但保留对它们自己的IR的识别特异性，是用于设计具有预测和改进的序列识别的嵌合转座酶的有前途的平台。
Most DNA transposons move from one genomic location to another by a cut-and-paste mechanism and are useful tools for genomic manipulations. Short inverted repeat (IR) DNA sequences marking each end of the transposon are recognized by a DNA transposase (encoded by the transposon itself). This enzyme cleaves the transposon ends and integrates them at a new genomic location. We report here a comparison of the biophysical and biochemical properties of two closely related and active mariner/Tc1 family DNA transposases: Mboumar-9 and Mos1. We compared the in vitro cleavage activities of the enzymes on their own IR sequences, as well as cross-recognition of their inverted repeat sequences. We found that, like Mos1, untagged recombinant Mboumar-9 transposase is a dimer and forms a stable complex with inverted repeat DNA in the presence of Mg2+ ions. Mboumar-9 transposase cleaves its inverted repeat DNA in the manner observed for Mos1 transposase. There was minimal cross-recognition of IR sequences between Mos1 and Mboumar-9 transposases, despite these enzymes having 68% identical amino acid sequences. Transposases sharing common biophysical and biochemical properties, but retaining recognition specificity toward their own IR, are a promising platform for the design of chimeric transposases with predicted and improved sequence recognition.
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影响因子：
14.9
作者：
Clark KJ;Carlson DF;Leaver MJ;Foster LK;Fahrenkrug SC
通讯作者：
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影响因子：
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作者：
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通讯作者：
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作者：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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