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旋毛虫感染的肌细胞:核斑点区域中丰富的RNA聚合酶II与核抗原共定位。

Trichinella spiralis-infected muscle cells: abundant RNA polymerase II in nuclear speckle domains colocalizes with nuclear antigens.

作者信息

Yao C, Jasmer D P

机构信息

Department of Internal Medicine and Microbiology, College of Medicine, University of Iowa, Iowa City 52242, USA.

出版信息

Infect Immun. 2001 Jun;69(6):4065-71. doi: 10.1128/IAI.69.6.4065-4071.2001.

Abstract

Infection of mammalian skeletal muscle cells by Trichinella spiralis causes host nuclei to become polyploid (ca. 4N) and abnormally enlarged. It has been postulated that this enlargement reflects an infection-induced elevation of host transcription. Anthelmintic treatment of T. spiralis-infected rodents with mebendazole (MBZ) causes a reduction in the size of infected cell nuclei and a significant reduction in the total RNA content of individual infected muscle cells. A monoclonal antibody to the large subunit of RNA polymerase II (Pol II) was used here to assess the effects of infection on Pol II levels in isolated infected cell nuclei. Pol II was localized to speckle domains in isolated infected cell nuclei. Similar domains have been previously localized to sites of RNA synthesis or processing. When compared to the levels in nuclei from other, uninfected host cells, speckle-localized Pol II (SL-Pol II) levels were significantly elevated in infected cell nuclei by a mean of 3.9- to 6.8-fold. Nuclear antigens (NA) recognized by antibodies against T. spiralis localized to infected cell nuclei. By use of confocal microscopy, a subpopulation of NA was found colocalized with most speckle domains defined by Pol II. MBZ treatment of chronically infected mice, which depletes NA from infected cell nuclei, caused a significant depletion of SL-Pol II from infected cell nuclei. Control nuclei had a mean of 70% more SL-Pol II than MBZ-treated nuclei. The mean residual level of Pol II in these polyploid nuclei remained elevated by 120% over the level in 2N control nuclei. These observations may indicate two distinct effects of infection on Pol II levels in host cells.

摘要

旋毛虫感染哺乳动物骨骼肌细胞会导致宿主细胞核变成多倍体(约4N)并异常增大。据推测,这种增大反映了感染诱导的宿主转录水平升高。用甲苯咪唑(MBZ)对感染旋毛虫的啮齿动物进行驱虫治疗,会使感染细胞核的大小减小,单个感染肌肉细胞的总RNA含量显著降低。本文使用一种针对RNA聚合酶II(Pol II)大亚基的单克隆抗体来评估感染对分离的感染细胞核中Pol II水平的影响。Pol II定位于分离的感染细胞核中的斑点结构域。此前已将类似的结构域定位到RNA合成或加工位点。与来自其他未感染宿主细胞的细胞核中的水平相比,感染细胞核中斑点定位的Pol II(SL-Pol II)水平显著升高,平均升高3.9至6.8倍。抗旋毛虫抗体识别的核抗原(NA)定位于感染细胞核。通过共聚焦显微镜观察,发现一部分NA与由Pol II定义的大多数斑点结构域共定位。对慢性感染小鼠进行MBZ治疗,可使感染细胞核中的NA减少,同时导致感染细胞核中的SL-Pol II显著减少。对照细胞核中的SL-Pol II平均比经MBZ处理的细胞核多70%。这些多倍体细胞核中Pol II的平均残留水平仍比2N对照细胞核中的水平高120%。这些观察结果可能表明感染对宿主细胞中Pol II水平有两种不同的影响。

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