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Leukotriene D(4) triggers an association between gbetagamma subunits and phospholipase C-gamma1 in intestinal epithelial cells

Thodeti, C K ; Adolfsson, J ; Juhas, M LU and Sjölander, A LU (2000) In Journal of Biological Chemistry 275(13). p.53-9849
Abstract

The proinflammatory mediator leukotriene D(4) (LTD(4)) binds to the seven-transmembrane receptor CYSLT(1). Although this leukotriene plays an important biological role, its intracellular signaling pathways are only partly known. In previous experiments, we found that LTD(4) induced tyrosine phosphorylation and translocation of phospholipase (PLC)-gamma1 to a plasma membrane fraction in a human epithelial cell line (Int 407). In the present study, we further examined these signaling events and found that LTD(4) induced a rapid interaction between Gbetagamma subunits and PLC-gamma1; results obtained with GST fusion proteins of PLC-gamma1 suggest that this interaction is mediated via the pleckstrin homology domain of PLC-gamma1. Moreover,... (More)

The proinflammatory mediator leukotriene D(4) (LTD(4)) binds to the seven-transmembrane receptor CYSLT(1). Although this leukotriene plays an important biological role, its intracellular signaling pathways are only partly known. In previous experiments, we found that LTD(4) induced tyrosine phosphorylation and translocation of phospholipase (PLC)-gamma1 to a plasma membrane fraction in a human epithelial cell line (Int 407). In the present study, we further examined these signaling events and found that LTD(4) induced a rapid interaction between Gbetagamma subunits and PLC-gamma1; results obtained with GST fusion proteins of PLC-gamma1 suggest that this interaction is mediated via the pleckstrin homology domain of PLC-gamma1. Moreover, LTD(4) induced an increased association of c-Src with PLC-gamma1, and the selective Src family tyrosine kinase inhibitor PP1 blocked both LTD(4)-induced tyrosine phosphorylation of PLC-gamma1 and the association of PLC-gamma1 with Gbetagamma subunits. The relevance of these observations in intracellular calcium signaling was investigated by microinjecting cells with anti-Gbeta, anti-PLC-gamma1, or anti-c-Src antibodies and by pretreatment with PP1. LTD(4)-induced calcium mobilization was blocked by each of the indicated antibodies (but not isotype-matched control antibodies) and by PP1. Our data suggest that Gbetagamma subunits can, directly or indirectly, serve as membrane-bound partners for PLC-gamma1 and c-Src and that each of these proteins is essential for LTD(4)-induced downstream PLC-gamma1 signaling.

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keywords
Calcium/metabolism, Calcium Signaling, Cells, Cultured, Epithelial Cells/cytology, GTP-Binding Proteins/metabolism, Humans, Intestinal Mucosa/metabolism, Isoenzymes/metabolism, Leukotriene D4/metabolism, Phospholipase C gamma, Protein Binding, Proto-Oncogene Proteins pp60(c-src)/metabolism, Type C Phospholipases/metabolism
in
Journal of Biological Chemistry
volume
275
issue
13
pages
5 pages
publisher
ASBMB
external identifiers
  • scopus:0034737450
  • pmid:10734140
ISSN
0021-9258
DOI
10.1074/jbc.275.13.9849
language
English
LU publication?
yes
id
669f6e65-d319-4c2f-8ede-fa9b64101441
date added to LUP
2019-07-03 11:27:05
date last changed
2021-08-04 01:21:25
@article{669f6e65-d319-4c2f-8ede-fa9b64101441,
  abstract     = {<p>The proinflammatory mediator leukotriene D(4) (LTD(4)) binds to the seven-transmembrane receptor CYSLT(1). Although this leukotriene plays an important biological role, its intracellular signaling pathways are only partly known. In previous experiments, we found that LTD(4) induced tyrosine phosphorylation and translocation of phospholipase (PLC)-gamma1 to a plasma membrane fraction in a human epithelial cell line (Int 407). In the present study, we further examined these signaling events and found that LTD(4) induced a rapid interaction between Gbetagamma subunits and PLC-gamma1; results obtained with GST fusion proteins of PLC-gamma1 suggest that this interaction is mediated via the pleckstrin homology domain of PLC-gamma1. Moreover, LTD(4) induced an increased association of c-Src with PLC-gamma1, and the selective Src family tyrosine kinase inhibitor PP1 blocked both LTD(4)-induced tyrosine phosphorylation of PLC-gamma1 and the association of PLC-gamma1 with Gbetagamma subunits. The relevance of these observations in intracellular calcium signaling was investigated by microinjecting cells with anti-Gbeta, anti-PLC-gamma1, or anti-c-Src antibodies and by pretreatment with PP1. LTD(4)-induced calcium mobilization was blocked by each of the indicated antibodies (but not isotype-matched control antibodies) and by PP1. Our data suggest that Gbetagamma subunits can, directly or indirectly, serve as membrane-bound partners for PLC-gamma1 and c-Src and that each of these proteins is essential for LTD(4)-induced downstream PLC-gamma1 signaling.</p>},
  author       = {Thodeti, C K and Adolfsson, J and Juhas, M and Sjölander, A},
  issn         = {0021-9258},
  language     = {eng},
  month        = {03},
  number       = {13},
  pages        = {53--9849},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Leukotriene D(4) triggers an association between gbetagamma subunits and phospholipase C-gamma1 in intestinal epithelial cells},
  url          = {http://dx.doi.org/10.1074/jbc.275.13.9849},
  doi          = {10.1074/jbc.275.13.9849},
  volume       = {275},
  year         = {2000},
}