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引用次数: 1
摘要
小麦矮小病毒 Rep 结构域是一种 HUH 内切酶，参与滚圆复制。HUH 内切酶或 HUH 标签通过二价金属离子配位形成共价蛋白质-ssDNA 加合物，从而裂解特定的 ssDNA 序列并形成磷酪氨酸连接。这种蛋白质-ssDNA 融合可用于各种生物技术应用，如细胞成像、细胞条形码、DNA 引导的蛋白质定位以及 DNA 系链蛋白质的单分子操作。解决 Rep 结构域与 DNA 复合物的结构问题可以提供有关 HUH-tag 序列特异性的必要信息，并允许对蛋白质-DNA 相互作用进行合理的工程设计。在这里，我们用衍射到 1.24 Å 的晶体解出了 WDV Rep 结构域的 apo 状态结构。不过，所解决的结构代表着向解决蛋白质-ssDNA复合物迈出了一步。
Crystal structure of the wheat dwarf virus Rep domain
The wheat dwarf virus Rep domain is an HUH‐endonuclease and is involved in rolling‐circle replication. HUH‐endonucleases, or HUH‐tags, form covalent protein‐ssDNA adducts by coordinating a divalent metal ion to cleave a specific ssDNA sequence and form a phosphotyrosine linkage. This protein‐ssDNA fusion is useful for various biotechnology applications such as cellular imaging, cellular barcoding, DNA‐guided protein localization, and single molecule manipulation of DNA‐tethered proteins. Solving the structure of the Rep domain in complex with DNA could present necessary information regarding HUH‐tag sequence specificity and allow for rational engineering of protein‐DNA interactions. Here, the structure of WDV Rep domain in the apo state was solved with a crystal diffracting to 1.24 Å. While ssDNA soaks were attempted, they proved ineffective. However, the solved structure represents a step towards solving protein‐ssDNA complex.
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