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Cj1496c Encodes a Campylobacter jejuni Glycoprotein That Influences Invasion of Human Epithelial Cells and Colonization of the Chick Gastrointestinal Tract

机译:Cj1496c编码空肠弯曲菌糖蛋白,它影响人上皮细胞的侵袭和小鸡胃肠道的定植

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摘要

Campylobacter jejuni has an N-linked protein glycosylation pathway that is required for efficient cell invasion and chick gastrointestinal colonization by the microbe. In this study, we constructed insertion mutants of 22 putative glycoprotein genes and examined the ability of each to invade the human intestinal epithelial cell line INT-407. Among the mutants tested, one carrying an insertion in Cj1496c was defective for invasion into INT-407 cells; this defect was also observed in an in-frame deletion mutant of Cj1496c (ΔCj1496c). The ΔCj1496c mutant C. jejuni also showed a reduced ability to colonize chick ceca. Site-specific mutagenesis combined with Western blot analysis suggested that the Cj1496c protein is glycosylated at N73 and N169. However, the ΔCj1496c mutant expressing a nonglycosylated form of Cj1496c exhibited levels of invasion and colonization equivalent to those of the parent strain, suggesting that glycans are not directly involved in the function of Cj1496c.
机译:空肠弯曲杆菌具有N-连接的蛋白质糖基化途径,该途径是微生物有效进行细胞侵袭和鸡胃肠道定殖所必需的。在这项研究中,我们构建了22个推定的糖蛋白基因的插入突变体,并检查了每个突变体侵袭人类肠道上皮细胞系INT-407的能力。在测试的突变体中,一个在Cj1496c中带有插入片段的突变体无法侵入INT-407细胞。在Cj1496c的框内缺失突变体(ΔCj1496c)中也观察到此缺陷。 ΔCj1496c突变空肠弯曲菌也显示出降低了对小盲肠定殖的能力。位点特异性诱变结合蛋白质印迹分析表明,Cj1496c蛋白在N73和N169处被糖基化。但是,表达非糖基化形式的Cj1496c的ΔCj1496c突变体表现出与亲本菌株相同的侵袭和定植水平,这表明聚糖并不直接参与Cj1496c的功能。

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