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Real-time visualization of PH domain-dependent translocation of phospholipase C-delta1 in renal epithelial cells (MDCK): response to hypo-osmotic stress.
Biochem Biophys Res Commun. 1999 Jan 19; 254(2):284-91.BB

Abstract

Green fluorescent protein (GFP)-tagged phospholipase C (PLC)-delta1 and its mutants were expressed in Madin-Darby canine kidney (MDCK) cells. GFP-PLC-delta1 or the GFP-tagged pleckstrin homology (PH) domain of PLC-delta1 itself was found to be predominantly localized at the plasma membrane. The DeltaPH mutant or a site-directed mutant containing a PH domain which does not bind inositol 1,4, 5-trisphosphate and cannot hydrolyze phosphatidylinositol 4, 5-bisphosphate in vitro was seen only in the cytosol. In living MDCK cells hypo-osmotic stress caused a rapid dissociation of GFP-PLC-delta1 from the plasma membrane, which coincided with phosphoinositide breakdown. A PLC inhibitor, U73122, blocked this translocation, but depletion of extracellular Ca2+ had no effect. The translocation was reversed by replacement with an iso-osmotic buffer. Our results demonstrate that the PH domain plays a critical role in the membrane targeting of PLC-delta1 and that the intracellular distribution of the enzyme is regulated by osmotic stress-driven phosphoinositide turnover.

Authors+Show Affiliations

Department of Life Science, Faculty of Science, Harima Science Garden City, Hyogo, 678-1297, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9918830

Citation

Fujii, M, et al. "Real-time Visualization of PH Domain-dependent Translocation of Phospholipase C-delta1 in Renal Epithelial Cells (MDCK): Response to Hypo-osmotic Stress." Biochemical and Biophysical Research Communications, vol. 254, no. 2, 1999, pp. 284-91.
Fujii M, Ohtsubo M, Ogawa T, et al. Real-time visualization of PH domain-dependent translocation of phospholipase C-delta1 in renal epithelial cells (MDCK): response to hypo-osmotic stress. Biochem Biophys Res Commun. 1999;254(2):284-91.
Fujii, M., Ohtsubo, M., Ogawa, T., Kamata, H., Hirata, H., & Yagisawa, H. (1999). Real-time visualization of PH domain-dependent translocation of phospholipase C-delta1 in renal epithelial cells (MDCK): response to hypo-osmotic stress. Biochemical and Biophysical Research Communications, 254(2), 284-91.
Fujii M, et al. Real-time Visualization of PH Domain-dependent Translocation of Phospholipase C-delta1 in Renal Epithelial Cells (MDCK): Response to Hypo-osmotic Stress. Biochem Biophys Res Commun. 1999 Jan 19;254(2):284-91. PubMed PMID: 9918830.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Real-time visualization of PH domain-dependent translocation of phospholipase C-delta1 in renal epithelial cells (MDCK): response to hypo-osmotic stress. AU - Fujii,M, AU - Ohtsubo,M, AU - Ogawa,T, AU - Kamata,H, AU - Hirata,H, AU - Yagisawa,H, PY - 1999/1/27/pubmed PY - 1999/1/27/medline PY - 1999/1/27/entrez SP - 284 EP - 91 JF - Biochemical and biophysical research communications JO - Biochem. Biophys. Res. Commun. VL - 254 IS - 2 N2 - Green fluorescent protein (GFP)-tagged phospholipase C (PLC)-delta1 and its mutants were expressed in Madin-Darby canine kidney (MDCK) cells. GFP-PLC-delta1 or the GFP-tagged pleckstrin homology (PH) domain of PLC-delta1 itself was found to be predominantly localized at the plasma membrane. The DeltaPH mutant or a site-directed mutant containing a PH domain which does not bind inositol 1,4, 5-trisphosphate and cannot hydrolyze phosphatidylinositol 4, 5-bisphosphate in vitro was seen only in the cytosol. In living MDCK cells hypo-osmotic stress caused a rapid dissociation of GFP-PLC-delta1 from the plasma membrane, which coincided with phosphoinositide breakdown. A PLC inhibitor, U73122, blocked this translocation, but depletion of extracellular Ca2+ had no effect. The translocation was reversed by replacement with an iso-osmotic buffer. Our results demonstrate that the PH domain plays a critical role in the membrane targeting of PLC-delta1 and that the intracellular distribution of the enzyme is regulated by osmotic stress-driven phosphoinositide turnover. SN - 0006-291X UR - https://www.unboundmedicine.com/medline/citation/9918830/Real_time_visualization_of_PH_domain_dependent_translocation_of_phospholipase_C_delta1_in_renal_epithelial_cells__MDCK_:_response_to_hypo_osmotic_stress_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-291X(98)99936-3 DB - PRIME DP - Unbound Medicine ER -