Yersinia enterocolitica can deliver Yop proteins into a wide range of cell types:: Development of a delivery system for heterologous proteins
Yersinia enterocolitica can deliver Yop proteins into a wide range of cell types:: Development of a delivery system for heterologous proteins
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DOI：
10.1078/0171-9335-00098
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发表时间：
2000-10-01
影响因子：
6.6
通讯作者：
Cornelis, GR
中科院分区：
文献类型：
--
作者：
Boyd, AP;Grosdent, N;Cornelis, GR
小肠结肠炎耶尔森氏菌将称为 Yop 效应子的毒力蛋白易位到真核细胞的细胞质中。在这里我们调查是否 Y;小肠结肠炎可以将 Yops 转移到一系列真核细胞中，包括神经元和昆虫细胞。是；小肠结肠炎将杂合报告蛋白 YopE-Cya 转移到每种测试的真核细胞类型中。此外，小肠结肠炎耶尔森氏菌对每种贴壁细胞类型都具有细胞毒性。因此，我们检测到小肠结肠炎耶尔森氏菌可以将Yops易位到的真核细胞范围没有限制，Yop效应子YopE、YopH和YopT对于测试的贴壁细胞类型均具有细胞毒性，这表明I：小肠结肠炎耶尔森氏菌不仅在将特定Yop效应子易位到每种细胞类型中没有选择性，而且这些Yop效应子的作用不是细胞类型特异性的。将 Yop 转位到非吞噬细胞中需要 Invasin 和/或 YadA 这两种强大的粘附素，但转位到巨噬细胞中则不需要。为了将耶尔森氏菌易位系统用于广泛的应用，构建了用于将异源蛋白递送至真核细胞的I：小肠结肠炎易位菌株和载体。这种菌株+载体组合缺乏易位的Yop效应子，并允许将与YopE的最小N端分泌/易位信号融合的异源蛋白递送到真核细胞中，使用这种策略将YopE-白喉毒素亚基易位混合蛋白进入多种细胞类型。
Y. enterocolitica translocates virulence proteins, called Yop effecters, into the cytosol of eukaryotic cells. Here we investigated whether Y; enterocolitica could translocate Yops into a range of eukaryotic cells including neurons and insect cells. Y; enterocolitica translocated the hybrid reporter protein YopE-Cya into each of the eukaryotic cell types tested. In addition, Y. enterocolitica was cytotoxic for each of the adherent cell types. Thus we detected no limit to the range of eukaryotic cells into which Y. enterocolitica can translocate Yops, The Yop effecters YopE, YopH and YopT were each cytotoxic for the adherent cell types tested, showing that not only is I: enterocolitica not selective in its translocation of particular Yop effecters into each cell type, but also that the action of these Yop effecters is not cell type specific. Invasin and/or YadA, two powerful adhesins were required for translocation of Yop into non-phagocytic cells but not for translocation into macrophages. To use the Yersinia translocation system for broad applications, a I: enterocolitica translocation strain and vector for the delivery of heterologous proteins into eukaryotic cells was constructed. This strain + vector combination lacks the translocated Yop effecters and allows delivery into eukaryotic cells of heterologous proteins fused to the minimal N-terminal secretion/translocation signal of YopE, Using this strategy translocation of a YopE-Diphtheria toxin subunit A hybrid protein into several cell types has been shown.