A protein kinase A-dependent mechanism by which rotavirus affects the distribution and mRNA level of the functional tight junction-associated protein, occludin, in human differentiated intestinal caco-2 cells
A protein kinase A-dependent mechanism by which rotavirus affects the distribution and mRNA level of the functional tight junction-associated protein, occludin, in human differentiated intestinal caco-2 cells
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DOI：
10.1128/jvi.00263-07
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发表时间：
2007-08-01
影响因子：
5.4
通讯作者：
Servin, Alain L.
中科院分区：
文献类型：
--
作者：
Beau, Isabelle;Cotte-Laffitte, Jacqueline;Servin, Alain L.
我们发现，在Caco-2细胞单层的紧密连接（TJs）处，恒河猴轮状病毒（RRV）感染导致occludin消失。共聚焦激光扫描显微镜显示occludin从细胞边界消失，但未改变其他tj相关蛋白ZO-1和ZO-3的表达。对rrv感染细胞的Western免疫印迹分析显示，Triton x -100不溶性和Triton x -100可溶部分中非磷酸化形式的occludin水平显著下降，磷酸化形式的occludin水平没有任何变化。定量逆转录pcr显示，在rrv感染的细胞中，编码occludin的基因的转录水平显著降低。在感染期间用rp -环AMP和蛋白激酶A抑制剂H89和KT5720处理rrv感染的细胞恢复了occludin的分布和编码occludin的基因的正常转录水平。
We found that at the tight junctions (TJs) of Caco-2 cell monolayers, rhesus monkey rotavirus (RRV) infection induced the disappearance of occludin. Confocal laser scanning microscopy showed the disappearance of occludin from the cell-cell boundaries without modifying the expression of the other TJ-associated proteins, ZO-1 and ZO-3. Western immunoblot analysis of RRV-infected cells showed a significant fall in the levels of the nonphosphorylated form of occludin in both Triton X-100-insoluble and Triton X-100-soluble fractions, without any change in the levels of the phosphorylated form of occludin. Quantitative reverse transcription-PCRs revealed that the level of transcription of the gene that encodes occludin was significantly reduced in RRV-infected cells. Treatment of RRV-infected cells with Rp-cyclic AMP and protein kinase A inhibitors H89 and KT5720 during the time course of the infection restored the distribution of occludin and a normal level of transcription of the gene that encodes occludin.