PMID- 12494451
OWN - NLM
STAT- MEDLINE
DCOM- 20030130
LR  - 20120625
IS  - 0021-9541 (Print)
IS  - 0021-9541 (Linking)
VI  - 194
IP  - 2
DP  - 2003 Feb
TI  - Galphaq signaling is required for Rho-dependent transcriptional activation of the 
      cyclooxygenase-2 promoter in fibroblasts.
PG  - 127-38
AB  - Previously, we demonstrated that the gastrin releasing peptide (GRP) induces 
      cyclooxygenase-2 (COX-2) expression through a Rho-dependent, protein kinase C 
      (PKC)-independent signaling pathway in fibroblasts (Slice et al., 1999, J Biol 
      Chem 274:27562-27566). However, the specific role of heterotrimeric guanine 
      nucleotide binding regulatory proteins (G-proteins) that are coupled to the GRP 
      receptor in Rho-dependent COX-2 expression has not been elucidated. In this 
      report, we utilize embryonic fibroblasts from transgenic mice containing double 
      gene knock-outs (DKO) for Galpha(q/11) and Galpha(12/13) to demonstrate that 
      COX-2 promoter activation by GRP requires Galpha(q). Furthermore, we show that 
      GRP-dependent COX-2 gene expression, as assessed by a COX-2 reporter luciferase 
      assay, was induced in cells lacking Galpha(12/13) but was blocked in cells that 
      did not express Galpha(q/11). GRP-dependent COX-2 promoter induction in 
      Galpha(q/11) deficient cells was rescued by expression of wild type Galpha(q) but 
      blocked by inhibition of calcium signaling in calcium-free media or in cells 
      treated with 2-aminoethoxydiphenylborate (2-APB). Co-stimulation of transfected 
      Galpha(q/11) deficient cells with GRP and thapsigargin (TG) induced the COX-2 
      promoter. Activation of endogenous Rho by expression of Onco-lbc or expression of 
      Rho A Q63L resulted in COX-2 promoter activation in Galpha(q/11) deficient cells. 
      Inhibition of Rho by Clostridium botulinum C3 toxin blocked COX-2 promoter 
      induction. Expression of Galpha(q) Q209L in the well-characterized fibroblast 
      cell line, NIH3T3, induced the COX-2 promoter which was blocked by expression of 
      C3 toxin. These results demonstrate that calcium signaling mediated by Galpha(q) 
      and Rho play critical roles in GRP-dependent COX-2 expression in fibroblasts.
CI  - Copyright 2002 Wiley-Liss, Inc.
FAU - Slice, Lee W
AU  - Slice LW
AD  - Division of Digestive Diseases, Department of Medicine, CURE: Digestive Diseases 
      Research Center, Greater Los Angeles VA Medical Center, University of California, 
      Los Angeles, California 90095, USA. lslice@mednet.ucla.edu
FAU - Han, Sang-Kyou
AU  - Han SK
FAU - Simon, Melvin I
AU  - Simon MI
LA  - eng
GR  - DK35740/DK/NIDDK NIH HHS/United States
GR  - GM-34236/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Cell Physiol
JT  - Journal of cellular physiology
JID - 0050222
RN  - 0 (Activating Transcription Factors)
RN  - 0 (Blood Proteins)
RN  - 0 (Eye Proteins)
RN  - 0 (Isoenzymes)
RN  - 0 (Protein Isoforms)
RN  - 0 (RNA, Messenger)
RN  - 0 (Rho Factor)
RN  - 0 (Transcription Factors)
RN  - 80043-53-4 (Gastrin-Releasing Peptide)
RN  - EC 1.14.99.1 (Cyclooxygenase 2)
RN  - EC 1.14.99.1 (Prostaglandin-Endoperoxide Synthases)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - EC 2.7.11.14 (G-Protein-Coupled Receptor Kinase 1)
RN  - EC 2.7.11.14 (Grk1 protein, mouse)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
RN  - EC 3.6.5.1 (GTP-Binding Protein alpha Subunits, Gi-Go)
RN  - EC 3.6.5.1 (GTP-Binding Protein alpha Subunits, Gq-G11)
RN  - EC 3.6.5.1 (Heterotrimeric GTP-Binding Proteins)
SB  - IM
MH  - Activating Transcription Factors
MH  - Animals
MH  - Blood Proteins/physiology
MH  - Calcium Signaling/physiology
MH  - Cyclooxygenase 2
MH  - *Eye Proteins
MH  - Fibroblasts/*physiology
MH  - G-Protein-Coupled Receptor Kinase 1
MH  - GTP-Binding Protein alpha Subunits, Gi-Go/physiology
MH  - GTP-Binding Protein alpha Subunits, Gq-G11
MH  - GTP-Binding Proteins/genetics
MH  - Gastrin-Releasing Peptide/physiology
MH  - Heterotrimeric GTP-Binding Proteins/genetics/*physiology
MH  - Isoenzymes/*genetics
MH  - Mice
MH  - Mice, Knockout
MH  - Promoter Regions, Genetic/*physiology
MH  - Prostaglandin-Endoperoxide Synthases/*genetics
MH  - Protein Isoforms/metabolism
MH  - Protein Kinase C/physiology
MH  - Protein Kinases/deficiency/genetics
MH  - RNA, Messenger/metabolism
MH  - Rho Factor/*physiology
MH  - Signal Transduction/*physiology
MH  - Transcription Factors/physiology
MH  - Transcriptional Activation/*physiology
EDAT- 2002/12/21 04:00
MHDA- 2003/01/31 04:00
CRDT- 2002/12/21 04:00
PHST- 2002/12/21 04:00 [pubmed]
PHST- 2003/01/31 04:00 [medline]
PHST- 2002/12/21 04:00 [entrez]
AID - 10.1002/jcp.10195 [doi]
PST - ppublish
SO  - J Cell Physiol. 2003 Feb;194(2):127-38. doi: 10.1002/jcp.10195.