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Mitogen activated protein kinases (MAPK) pathways regulate subcellular localization of claudins ‐6, ‐7 and ‐9 in transfected LLC‐PK1 cells
Author(s) -
ZavalaZendejas V E,
CastañedaPatlán C,
RendónHuerta E P
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a191
Subject(s) - claudin , transfection , tight junction , subcellular localization , immunoprecipitation , microbiology and biotechnology , mapk/erk pathway , cytosol , biology , kinase , chemistry , cell culture , cytoplasm , biochemistry , enzyme , genetics
Claudin family proteins play an important role in the barrier function of tight junctions (TJ), preserving cell polarity and determining the size and charge of the molecules that pass trough the paracelular space in epithelial and endothelial tissues. Previous studies demonstrate overexpression of several claudins in different neoplastic tissues. Our group found not only overexpression of claudins ‐6, ‐7 and ‐9, but also subcellular redistribution of these proteins in gastric adenocarcinoma. Our objective was to determine the role of the MAPK pathways in the regulation of the subcellular localization of claudins 6, 7 and 9 in LLC‐PK 1 cells. The cells were transfected with plasmids encoding each of these claudins tagged with GFP at their NH 2 terminus. Stable transfectants were treated with TPA and several MAPK inhibitors. Transepithelial electrical resistance (TER) was measured and the cells were observed by confocal microscopy. Immunoprecipitation was made in order to detect interaction of these claudins with other TJ proteins. TPA treatment induced a significant TER decrease in claudin‐transfected cells, whereas PD98059 inhibited the TER change. The GFP tagged claudins moved from membrane to cytosol when the cells were treated with TPA. This change in subcellular localization could be blocked with the PD98059 inhibitor. Supported by DGAPA IN225106