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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Domain activities of PapC usher reveal the mechanism of action of an Escherichia coli molecular machine
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Domain activities of PapC usher reveal the mechanism of action of an Escherichia coli molecular machine

机译：PapC的域活动揭示了大肠杆菌分子机器的作用机理

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摘要

P pili are prototypical chaperone-usher pathway-assembled pili used by Gram-negative bacteria to adhere to host tissues. The PapC usher contains five functional domains: a transmembrane (S-barrel, a p-sandwich Plug, an N-terminal (periplasmic) domain (NTD), and two C-terminal (periplasmic) domains, CTD1 and CTD2. Here, we delineated usher domain interactions between themselves and with chaperone-subunit complexes and showed that overexpres-sion of individual usher domains inhibits pilus assembly. Prior work revealed that the Plug domain occludes the pore of the transmembrane domain of a solitary usher, but the chaperone-adhesin-bound usher has its Plug displaced from the pore, adjacent to the NTD. We demonstrate an interaction between the NTD and Plug domains that suggests a biophysical basis for usher gating. Furthermore, we found that the NTD exhibits high-affinity binding to the chaperone-adhesin (PapDG) complex and low-affinity binding to the major tip subunit PapE (PapDE). We also demonstrate that CTD2 binds with lower affinity to all tested chaperone-subunit complexes except for the chaperone-terminator subunit (PapDH) and has a catalytic role in dissociating the NTD-PapDG complex, suggesting an interplay between recruitment to the NTD and transfer to CTD2 during pilus initiation. The Plug domain and the NTD-Plug complex bound all of the chaperone-subunit complexes tested including PapDH, suggesting that the Plug actively recruits chaperone-subunit complexes to the usher and is the sole recruiter of PapDH. Overall, our studies reveal the cooperative, active roles played by periplasmic domains of the usher to initiate, grow, and terminate a prototypical chaperone-usher pathway pilus.
机译：P菌毛是革兰氏阴性细菌用来粘附宿主组织的典型伴侣-引导途径组装菌毛。 PapC引入程序包含五个功能域：一个跨膜（S桶，一个p三明治塞子，一个N端（周质）域（NTD）和两个C端（周质）域CTD1和CTD2。描绘了它们之间以及与伴侣-亚基复合物之间的引入域相互作用，并表明单个引入域的过度表达会抑制菌毛的组装；先前的研究表明，Plug域阻塞了单个引入域的跨膜结构域的孔，但是伴侣-粘附素结合的引诱剂的Plug从孔中移出，与NTD相邻，我们证明了NTD和Plug域之间的相互作用为引诱选通提供了生物物理基础，此外，我们发现NTD与伴侣分子具有高亲和力结合。 -粘附素（PapDG）复合物和低亲和力与主要末端亚基PapE（PapDE）结合我们还证明CTD2与所有受试伴侣-亚基复合物的结合亲和力较低，除了伴侣-末端Nator亚基（PapDH），并在解离NTD-PapDG复合物中起催化作用，提示菌毛启动过程中募集NTD和转移至CTD2之间存在相互作用。 Plug域和NTD-Plug复合物结合了所有测试的伴侣-亚基复合物，包括PapDH，这表明Plug积极招募伴侣-亚基复合物来迎接迎宾者，并且是PapDH的唯一招募者。总体而言，我们的研究揭示了引物的周质域在启动，生长和终止典型的伴侣-诱导途径纤毛中所起的协同，积极作用。

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  • 作者

    Ender Volkan; Bradley A.Ford; Jerome S.Pinkner; Karen W.Dodson; Nadine S.Henderson; David G.Thanassi; Gabriel Waksman; Scott J.Hultgren;

  • 作者单位

    Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110,Center for Women's Infectious Diseases Research, Washington University School of Medicine, St. Louis, MO 63110;

    Center for Women's Infectious Diseases Research, Washington University School of Medicine, St. Louis, MO 63110,Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110;

    Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110,Center for Women's Infectious Diseases Research, Washington University School of Medicine, St. Louis, MO 63110;

    Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110,Center for Women's Infectious Diseases Research, Washington University School of Medicine, St. Louis, MO 63110;

    Center for Infectious Diseases, Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794;

    Center for Infectious Diseases, Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794;

    Institute of Structural and Molecular Biology, Birkbeck College and University College London, London WC1E 7HX,United Kingdom;

    Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110,Center for Women's Infectious Diseases Research, Washington University School of Medicine, St. Louis, MO 63110;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    biolayer interferometry; macromolecular assembly; urinary tract; infections; virulence factor; bacterial pathogenesis;

    机译：生物层干涉仪大分子组装尿路;感染;毒力因子;细菌发病;

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