Swan David G, Stadler Laura, Okan Emel, Hoffs Michael, Katzer Frank, Kinnaird Jane, McKellar Sue, Shiels Brian R
Department of Veterinary Parasitology, University of Glasgow, Bearsden Road, Glasgow, G61 1QH, UK.
Cell Microbiol. 2003 Dec;5(12):947-56. doi: 10.1046/j.1462-5822.2003.00340.x.
The intracellular apicomplexan parasite, Theileria annulata, manipulates its bovine host cell by over-riding the cells natural apoptotic response and inducing proliferation of the infected leukocyte. We have recently identified a T. annulata encoded family of polypeptides (TashATs) with characteristics that indicate that they are involved in control of host cell gene expression. Here we present data on another member of this family, TashHN, showing that it is located to the parasite and host cell nucleus. Immunoblot analysis demonstrated that, unlike TashAT2 and 3, TashHN displays three forms, the largest of which is enriched in the host nuclear fraction and appears to be phosphorylated. Northern and 5 prime race analyses identified multiple TashHN RNA species in infected cells that have retained the ability to differentiate. These transcripts showed subtly different kinetics, but all decreased during differentiation to the merozite, and two showed reduced levels prior to down-regulation of the other TashATs. In addition, analyses of multiple cell lines that have become severely attenuated in their potential to differentiate, indicated a substantial increase in TashHN expression, with host nuclear reactivity particularly enhanced.
细胞内顶复门寄生虫环形泰勒虫通过抑制宿主细胞的自然凋亡反应并诱导被感染白细胞的增殖来操控其牛宿主细胞。我们最近鉴定出了一个环形泰勒虫编码的多肽家族(TashATs),其特征表明它们参与宿主细胞基因表达的调控。在此,我们展示了该家族另一个成员TashHN的数据,表明它定位于寄生虫和宿主细胞核。免疫印迹分析表明,与TashAT2和3不同,TashHN呈现三种形式,其中最大的一种在宿主核组分中富集且似乎被磷酸化。Northern印迹和5'端快速扩增cDNA末端分析在感染细胞中鉴定出多种保留分化能力的TashHN RNA种类。这些转录本显示出细微不同的动力学,但在分化为裂殖子时均减少,并且其中两种在其他TashATs下调之前水平就已降低。此外,对多个分化潜能严重减弱的细胞系的分析表明,TashHN表达大幅增加,宿主核反应性尤其增强。