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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Dissection of the Burkholderia intracellular life cycle using a photothermal nanoblade
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Dissection of the Burkholderia intracellular life cycle using a photothermal nanoblade

机译:使用光热纳米刀片解剖伯克霍尔德菌细胞内生命周期

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摘要

Burkholderia pseudomallei and Burkholderia thailandensis are related pathogens that invade a variety of cell types, replicate in the cytoplasm, and spread to nearby cells. We have investigated temporal and spatial requirements for virulence determinants in the intracellular life cycle, using genetic dissection and photo-thermal nanoblade delivery, which allows efficient placement of bacterium-sized cargo into the cytoplasm of mammalian cells. The conserved Bsa type III secretion system (T3SS_(Bsa)) is dispensable for invasion, but is essential for escape from primary endosomes. By nanoblade delivery of B. thailandensis we demonstrate that all subsequent events in intercellular spread occur independently of T3SS_(Bsa) activity. Although intracellular movement was essential for cell-cell spread by B. pseudomallei and B. thailandensis, neither BimA-mediated actin polymerization nor the formation of membrane protrusions containing bacteria was required for B. thailandensis. Surprisingly,the cryptic (fla2) flagellar system encoded on chromosome 2 of B. thailandensis supported rapid intracellular motility and efficient cell-cell spread. Plaque formation by both pathogens was dependent on the activity of a type VI secretion system (T6SS-1) that functions downstream from T3SS_(Bsa) -mediated endosome escape. A remarkable feature of Burkholderia is their ability to induce the formation of multinucleate giant cells (MNGCs) in multiple cell types. By infection and nanoblade delivery, we observed complete correspondence between mutant phenotypes in assays for cell fusion and plaque formation, and time-course studies showed that plaque formation represents MNGC death. Our data suggest that the primary means for intercellular spread involves cell fusion, as opposed to pseudopod engulfment and bacterial escape from double-membrane vacuoles.
机译:假伯克霍尔德氏菌和泰国伯克霍尔德氏菌是相关病原体,它们侵入多种细胞类型,在细胞质中复制,并扩散到附近的细胞中。我们已经研究了使用细胞内遗传切割和光热纳米叶片递送技术在细胞内生命周期中对毒性决定因素的时间和空间要求,这可以将细菌大小的货物有效地放置到哺乳动物细胞的细胞质中。保守的Bsa III型分泌系统(T3SS_(Bsa))对于入侵是必不可少的,但对于从初级内体逃逸是必不可少的。通过B. thailandensis的纳米叶片传递,我们证明了细胞间扩散的所有后续事件均独立于T3SS_(Bsa)活性而发生。尽管胞内运动对于假小芽孢杆菌和泰国芽孢杆菌的细胞扩散是必不可少的,但对于芽孢杆菌来说,BimA介导的肌动蛋白聚合反应或包含细菌的膜突起的形成都不需要。出乎意料的是,泰国芽孢杆菌2号染色体上编码的(fla2)鞭毛系统支持快速的细胞内运动性和有效的细胞扩散。两种病原体形成的斑块取决于VI型分泌系统(T6SS-1)的活性,该系统在T3SS_(Bsa)介导的内体逃逸的下游起作用。伯克霍尔德氏菌的显着特征是它们能够诱导多种细胞类型中多核巨细胞(MNGC)的形成。通过感染和纳米刀片传递,我们在细胞融合和噬斑形成的测定中观察到突变表型之间的完全对应,并且时程研究表明,噬斑形成代表MNGC死亡。我们的数据表明,细胞间扩散的主要手段涉及细胞融合,而不是假足荚膜吞噬和细菌从双膜液泡中逸出。

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    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095;

    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095;

    Department of Electrical Engineering, University of California, Los Angeles, CA 90095;

    Department of Pathology and Laboratory Medicine, University of California, Los Angeles, CA 90095;

    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095;

    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095;

    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095;

    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095;

    California NanoSystems Institute, University of California, Los Angeles, CA 90095 Department of Mechanical and Aerospace Engineering, University of California, Los Angeles, CA 90095;

    Department of Pathology and Laboratory Medicine, University of California, Los Angeles, CA 90095 California NanoSystems Institute, University of California, Los Angeles, CA 90095 Broad Stem Cell Research Center, University of California, Los Angeles, CA 90095 Jonsson Cancer Center, University of California, Los Angeles, CA 90095;

    Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095 California NanoSystems Institute, University of California, Los Angeles, CA 90095 Molecular Biology Institute, University of California, Los Angeles, CA 90095;

  • 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);
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