Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Caco-2 Cells
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Carbazoles / pharmacology
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Cell Fractionation
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Cyclic AMP / analogs & derivatives
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Cyclic AMP / pharmacology
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Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
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Cyclic AMP-Dependent Protein Kinases / metabolism*
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Humans
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Indoles / pharmacology
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Intestines / cytology
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Intestines / enzymology
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Intestines / virology*
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Isoquinolines / pharmacology
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Membrane Proteins / analysis
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Microscopy, Confocal
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Occludin
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Octoxynol / chemistry
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Phosphoproteins / analysis
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Phosphoproteins / metabolism
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Phosphorylation
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Protein Kinase Inhibitors / pharmacology
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Pyrroles / pharmacology
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RNA, Messenger / analysis
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RNA, Messenger / metabolism
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Rotavirus / physiology*
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Sulfonamides / pharmacology
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Thionucleotides / pharmacology
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Tight Junctions / chemistry
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Tight Junctions / enzymology
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Tight Junctions / virology*
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Transcription, Genetic
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Zonula Occludens-1 Protein
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Zonula Occludens-2 Protein
Substances
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Carbazoles
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Indoles
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Isoquinolines
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Membrane Proteins
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OCLN protein, human
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Occludin
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Phosphoproteins
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Protein Kinase Inhibitors
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Pyrroles
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RNA, Messenger
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Sulfonamides
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TJP1 protein, human
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TJP2 protein, human
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Thionucleotides
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Zonula Occludens-1 Protein
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Zonula Occludens-2 Protein
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adenosine-3',5'-cyclic phosphorothioate
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KT 5720
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Octoxynol
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Cyclic AMP
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Cyclic AMP-Dependent Protein Kinases
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N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide