LISTERIA-MONOCYTOGENES MOVES RAPIDLY THROUGH THE HOST-CELL CYTOPLASM BY INDUCING DIRECTIONAL ACTIN ASSEMBLY
LISTERIA-MONOCYTOGENES MOVES RAPIDLY THROUGH THE HOST-CELL CYTOPLASM BY INDUCING DIRECTIONAL ACTIN ASSEMBLY
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DOI：
10.1073/pnas.87.16.6068
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发表时间：
1990-08-01
影响因子：
11.1
通讯作者：
SOUTHWICK, FS
中科院分区：
文献类型：
--
作者：
DABIRI, GA;SANGER, JM;SOUTHWICK, FS
单核细胞增生李斯特菌是一种细胞内寄生虫，可以很容易地感染巨噬细胞样细胞系J774和肾上皮细胞PtK 2。在被摄入后，生物体从吞噬溶酶体逃逸到宿主细胞的细胞质中。N-（7-硝基苯-2-氧杂-1，3-二氮唑-4-基）-鬼笔毒肽（phallacidin）是肌动蛋白丝（F-肌动蛋白）的特异性染色剂，表明在感染开始后1小时内，细菌被宿主细胞质肌动蛋白丝包围。到3小时，F-肌动蛋白的长突起开始在细菌的一端形成。这些肌动蛋白结构与肌动蛋白捆绑蛋白α-肌动蛋白以及原肌球蛋白。显微注射荧光标记的α-活细胞中的肌动蛋白表明，这些F-肌动蛋白突起的形成与细菌运动有关，当肌动蛋白丝通过细胞质迁移时，它们在细菌后面快速组装。这些F-肌动蛋白尾的长度达到40 μ m。细菌通过细胞质的运动是快速的，0.12-1.46 μ m/sec。在细胞松弛素D（0.5 μ g/ml）处理的2分钟内，所有细菌细胞内运动停止，并且另外的细菌相关肌动蛋白组装被阻断。非运动性李斯特菌突变体诱导可比较的肌动蛋白组装，并以与野生型相似的速度移动，这表明细胞内细菌移动所需的力是由宿主细胞产生的。L.单核细胞增多症可以以定向方式显著刺激宿主细胞肌动蛋白组装，其用于快速推动细菌通过细胞质，允许生物体移动到外周膜并扩散到未感染的细胞。
Listeria monocytogenes is an intracellular parasite that can readily infect the macrophage-like cell line J774 and the kidney epithelial cell PtK2. After being ingested, the organism escapes from the phagolysosome into the host-cell cytoplasm. N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)-phallacidin, a specific stain for actin filaments (F-actin), demonstrates that within 1 hr of initiation of infection, the bacteria become surrounded by host-cell cytoplasmic actin filaments. By 3 hr, long projections of F-actin begin to form at one end of the bacteria. These actin structures colocalize with the actin-bundling protein .alpha.-actinin as well as with tropomyosin. Microinjection of fluorescently labeled .alpha.-actinin in living cells demonstrates that the formation of these F-actin projections is associated with bacterial movement, actin filaments rapidly assembling behind the bacteria as they migrate through the cytoplasm. These F-actin tails attain lengths up to 40 .mu.m. The movement of the bacteria through the cytoplasm is rapid, 0.12-1.46 .mu.m/sec. Within 2 min of cytochalasin D (0.5 .mu.g/ml) treatment, all bacterial intracellular movement stops, and additional bacteria-associated actin assembly is blocked. A nonmotile Listeria mutant induces comparable actin assembly and moves at speeds similar to the wild type, indicating that the forces required for intracellular bacterial movement are generated by the host cell. L. monocytogenes can dramatically stimulate host-cell actin assembly in a directional manner, which serves to rapidly propel the bacteria through the cytoplasm, allowing the organisms to move to peripheral membranes and spread to uninfected cells.