Calcium-sensing receptor activation of Rho involves filamin and rho-guanine nucleotide exchange factor

Min Pi, Robert F. Spurney, T. U. Qisheng, Todd Hinson, Leigh Quarles

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

We investigated the role of Gαq, filamin, Rho, the RhoGEF Lbc, and the C terminus of calcium-sensing receptor (CasR) in CasR signaling. We found that Ca2+, Mg2+, or the calcimimetic R isomer of N-(3-[2-chlorophenyl]propyl)-(R)-α-methyl-3-methoxybenzylamine (NPS-R568) stimulated serum response element (SRE) activity human embryonic kidney 293 cells transfected with CasR and an SRE-luciferase reporter construct. Coexpression of either the dominant negative Gαq(305-359) minigene, regulators of G protein signaling (RGS)2 or RGS4, inhibited CasR-stimulated SRE activity, consistent with CasR activation of Gαq. The cytoskeletal associated Rho protein is involved CasR activation of SRE, as evidenced by CasR-mediated increase in membrane-associated Rho A and by the ability of Clostridium botulinum C3 (C3) exoenzyme to inhibit both CasR and GαqQL-stimulated SRE activity. Overexpression of the RhoGEF Lbc, lacking either the Dbl-homology or Pleckstrin homology domain, as well as the filamin peptide (1530-1875) inhibited CasR-mediated activation of SRE. A carboxyl-terminal CasR minigene, CasR(906-980), encoding a filamin binding region, also blocked CasR- and GαqQL-stimulated SRE activity. Potential interactions between CasR, RhoGEF Lbc, Rho A, Gαq, and filamin were demonstrated by reciprocal coimmunoprecipitation studies. Our results suggest that the C terminus of CasR may interact with filamin to create a cytoskeletal scaffold necessary for the spatial organization of Gαq, RhoGEF Lbc, and Rho signaling pathways upstream of SRE activation.

Original languageEnglish (US)
Pages (from-to)3830-3838
Number of pages9
JournalEndocrinology
Volume143
Issue number10
DOIs
StatePublished - Oct 1 2002
Externally publishedYes

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Rho Guanine Nucleotide Exchange Factors
Filamins
Calcium-Sensing Receptors
Serum Response Element
N-(2-chlorophenylpropyl)-1-(3-methoxyphenyl)ethylamine
GTP-Binding Protein Regulators
Clostridium botulinum

All Science Journal Classification (ASJC) codes

  • Endocrinology

Cite this

Calcium-sensing receptor activation of Rho involves filamin and rho-guanine nucleotide exchange factor. / Pi, Min; Spurney, Robert F.; Qisheng, T. U.; Hinson, Todd; Quarles, Leigh.

In: Endocrinology, Vol. 143, No. 10, 01.10.2002, p. 3830-3838.

Research output: Contribution to journalArticle

Pi, Min ; Spurney, Robert F. ; Qisheng, T. U. ; Hinson, Todd ; Quarles, Leigh. / Calcium-sensing receptor activation of Rho involves filamin and rho-guanine nucleotide exchange factor. In: Endocrinology. 2002 ; Vol. 143, No. 10. pp. 3830-3838.
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AB - We investigated the role of Gαq, filamin, Rho, the RhoGEF Lbc, and the C terminus of calcium-sensing receptor (CasR) in CasR signaling. We found that Ca2+, Mg2+, or the calcimimetic R isomer of N-(3-[2-chlorophenyl]propyl)-(R)-α-methyl-3-methoxybenzylamine (NPS-R568) stimulated serum response element (SRE) activity human embryonic kidney 293 cells transfected with CasR and an SRE-luciferase reporter construct. Coexpression of either the dominant negative Gαq(305-359) minigene, regulators of G protein signaling (RGS)2 or RGS4, inhibited CasR-stimulated SRE activity, consistent with CasR activation of Gαq. The cytoskeletal associated Rho protein is involved CasR activation of SRE, as evidenced by CasR-mediated increase in membrane-associated Rho A and by the ability of Clostridium botulinum C3 (C3) exoenzyme to inhibit both CasR and GαqQL-stimulated SRE activity. Overexpression of the RhoGEF Lbc, lacking either the Dbl-homology or Pleckstrin homology domain, as well as the filamin peptide (1530-1875) inhibited CasR-mediated activation of SRE. A carboxyl-terminal CasR minigene, CasR(906-980), encoding a filamin binding region, also blocked CasR- and GαqQL-stimulated SRE activity. Potential interactions between CasR, RhoGEF Lbc, Rho A, Gαq, and filamin were demonstrated by reciprocal coimmunoprecipitation studies. Our results suggest that the C terminus of CasR may interact with filamin to create a cytoskeletal scaffold necessary for the spatial organization of Gαq, RhoGEF Lbc, and Rho signaling pathways upstream of SRE activation.

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