NDLI: Involvement of Gαi2 in the Maintenance and Biogenesis of Epithelial Cell Tight Junctions

Content Provider	Journal of Biological Chemistry (JBC)
Author	Saha, Chandana Nigam, Sanjay K. Denker, Bradley M.
Abstract	Polarized epithelial cells have highly developed tight junctions (TJ) to maintain an impermeant barrier and segregate plasma membrane functions, but the mechanisms that promote TJ formation and maintain its integrity are only partially defined. Treatment of confluent monolayers of Madin-Darby canine kidney (MDCK) cells with AlF4 − (activator of heterotrimeric G protein α subunits) results in a 3–4-fold increase in transepithelial resistances (TER), a reliable indicator of TJ integrity. MOCK cells transfected with activated Gα0 (Q205L) have acclerated TJ formation (Denker, B. M., Saha, C., Khawaja, S., and Nigam, S. J. (1996) J. Biol. Chem. 271, 25750–25753). Gαi2 has been localized within the tight junction, and a role for Gαi2in the formation and/or maintenance of the tight junction was studied by transfection of MDCK cells with vector without insert (PC), wild type Gαi2, or a GTPase-deficient mutant (constitutively activated), Q205Lαi2. Tryptic conformational analysis confirmed expression of a constitutively active Gαi2 in Q205Lαi2-MDCK cells, and confocal microscopy showed a similar pattern of Gαi2 localization in the three cell lines. Q205Lαi2-MDCK cells had significantly higher base-line TER values than wild type Gαi2- or PC-MDCK cells (1187 ± 150 versus 576 ± 89 (Gαi2); 377 ± 52 Ω·cm2 (PC)), and both Gαi2- and Q205Lαi2-transfected cell lines more rapidly develop TER in the Ca2+ switch, a model widely used to study the mechanisms of junctional assembly. Treatment of cells with AlF4 − during the Ca2+ switch had little effect on the kinetics of TER development in Gαi2- or Q205Lαi2-MDCK cells, but PC cells reached half-maximal TER significantly sooner in the presence of AlF4 − (similar times to Gαi2-transfected cells). Base-line TER values obtained after the switch were significantly higher for all three cell lines in the presence of AlF4 −. These findings indicate that Gαi2 is important for both the maintenance and development of the TJ, although additional Gα subunits are likely to play a role.
Related Links	http://www.jbc.org/content/273/34/21629.abstract
Ending Page	21633
Starting Page	21629
Page Count	5
File Format	HTM / HTML PDF
ISSN	00219258
Journal	Journal of Biological Chemistry (JBC)
Issue Number	34
Volume Number	273
DOI	10.1074/jbc.273.34.21629
e-ISSN	1083351X
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	1998-08-21
Access Restriction	Open
Subject Keyword	Tight junction (TJ) Madin-Darby canine kidney (MDCK) Guanine nucleotide-binding protein (G protein) Zona occludens (ZO) Transepithelial resistance (TER) Protein kinase C (PKC) Normal calcium (NC) Phosphate-buffered saline (PBS) Guanosine 5′-3-O-(thio)triphosphate. (GTPγS) CELL BIOLOGY AND METABOLISM
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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