Constitutive Activation of NF-κB and Secretion of Interleukin-8 Induced by the G Protein-coupled Receptor of Kaposi's Sarcoma-associated Herpesvirus Involve Gα13 and RhoA

Larry W. Shepard, Ming Yang, Ping Xie, Darren D Browning, Tatyana Voyno-Yasenetskaya, Tohru Kozasa, Richard D. Ye

Research output: Contribution to journalArticle

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Abstract

The Kaposi's sarcoma herpesvirus (KSHV) open reading frame 74 encodes a G protein-coupled receptor (GPCR) for chemokines. Exogenous expression of this constitutively active GPCR leads to cell transformation and vascular overgrowth characteristic of Kaposi's sarcoma. We show here that expression of KSHV-GPCR in transfected cells results in constitutive transactivation of nuclear factor κB (NF-κB) and secretion of interleukin-8, and this response involves activation of Gα13 and RhoA. The induced expression of a NF-κB luciferase reporter was partially reduced by pertussis toxin and the Gβγ scavenger transducin, and enhanced by co-expression of Gα13 and to a lesser extent, Gαq. These results indicate coupling of KSHV-GPCR to multiple G proteins for NF-κB activation. Expression of KSHV-GPCR led to stress fiber formation in NIH 3T3 cells. To examine the involvement of the Gα13-RhoA pathway in KSHV-GPCR-mediated NF-κB activation, HeLa cells were transfected with KSHV-GPCR alone and in combination with the regulator of G protein signaling (RGS) from p115RhoGEF or a dominant negative RhoA(T19N). Both constructs, as well as the C3 exoenzyme from Clostritium botulinum, partially reduced NF-κB activation by KSHV-GPCR, and by a constitutively active Gα13(Q226L). KSHV-GPCR-induced NF-κB activation is accompanied by increased secretion of IL-8, a function mimicked by the activated Gα13 but not by an activated Gα q(Q209L). These results suggest coupling of KSHV-GPCR to the Gα13-RhoA pathway in addition to other G proteins.

Original languageEnglish (US)
Pages (from-to)45979-45987
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number49
DOIs
StatePublished - Dec 7 2001
Externally publishedYes

Fingerprint

Human Herpesvirus 8
Kaposi's Sarcoma
G-Protein-Coupled Receptors
Herpesviridae
Interleukin-8
Chemical activation
GTP-Binding Proteins
Rho Guanine Nucleotide Exchange Factors
GTP-Binding Protein Regulators
Transducin
NIH 3T3 Cells
Stress Fibers
Pertussis Toxin
Luciferases
HeLa Cells
Chemokines
Transcriptional Activation
Open Reading Frames
Blood Vessels

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Constitutive Activation of NF-κB and Secretion of Interleukin-8 Induced by the G Protein-coupled Receptor of Kaposi's Sarcoma-associated Herpesvirus Involve Gα13 and RhoA. / Shepard, Larry W.; Yang, Ming; Xie, Ping; Browning, Darren D; Voyno-Yasenetskaya, Tatyana; Kozasa, Tohru; Ye, Richard D.

In: Journal of Biological Chemistry, Vol. 276, No. 49, 07.12.2001, p. 45979-45987.

Research output: Contribution to journalArticle

Shepard, Larry W. ; Yang, Ming ; Xie, Ping ; Browning, Darren D ; Voyno-Yasenetskaya, Tatyana ; Kozasa, Tohru ; Ye, Richard D. / Constitutive Activation of NF-κB and Secretion of Interleukin-8 Induced by the G Protein-coupled Receptor of Kaposi's Sarcoma-associated Herpesvirus Involve Gα13 and RhoA. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 49. pp. 45979-45987.
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abstract = "The Kaposi's sarcoma herpesvirus (KSHV) open reading frame 74 encodes a G protein-coupled receptor (GPCR) for chemokines. Exogenous expression of this constitutively active GPCR leads to cell transformation and vascular overgrowth characteristic of Kaposi's sarcoma. We show here that expression of KSHV-GPCR in transfected cells results in constitutive transactivation of nuclear factor κB (NF-κB) and secretion of interleukin-8, and this response involves activation of Gα13 and RhoA. The induced expression of a NF-κB luciferase reporter was partially reduced by pertussis toxin and the Gβγ scavenger transducin, and enhanced by co-expression of Gα13 and to a lesser extent, Gαq. These results indicate coupling of KSHV-GPCR to multiple G proteins for NF-κB activation. Expression of KSHV-GPCR led to stress fiber formation in NIH 3T3 cells. To examine the involvement of the Gα13-RhoA pathway in KSHV-GPCR-mediated NF-κB activation, HeLa cells were transfected with KSHV-GPCR alone and in combination with the regulator of G protein signaling (RGS) from p115RhoGEF or a dominant negative RhoA(T19N). Both constructs, as well as the C3 exoenzyme from Clostritium botulinum, partially reduced NF-κB activation by KSHV-GPCR, and by a constitutively active Gα13(Q226L). KSHV-GPCR-induced NF-κB activation is accompanied by increased secretion of IL-8, a function mimicked by the activated Gα13 but not by an activated Gα q(Q209L). These results suggest coupling of KSHV-GPCR to the Gα13-RhoA pathway in addition to other G proteins.",
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T1 - Constitutive Activation of NF-κB and Secretion of Interleukin-8 Induced by the G Protein-coupled Receptor of Kaposi's Sarcoma-associated Herpesvirus Involve Gα13 and RhoA

AU - Shepard, Larry W.

AU - Yang, Ming

AU - Xie, Ping

AU - Browning, Darren D

AU - Voyno-Yasenetskaya, Tatyana

AU - Kozasa, Tohru

AU - Ye, Richard D.

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AB - The Kaposi's sarcoma herpesvirus (KSHV) open reading frame 74 encodes a G protein-coupled receptor (GPCR) for chemokines. Exogenous expression of this constitutively active GPCR leads to cell transformation and vascular overgrowth characteristic of Kaposi's sarcoma. We show here that expression of KSHV-GPCR in transfected cells results in constitutive transactivation of nuclear factor κB (NF-κB) and secretion of interleukin-8, and this response involves activation of Gα13 and RhoA. The induced expression of a NF-κB luciferase reporter was partially reduced by pertussis toxin and the Gβγ scavenger transducin, and enhanced by co-expression of Gα13 and to a lesser extent, Gαq. These results indicate coupling of KSHV-GPCR to multiple G proteins for NF-κB activation. Expression of KSHV-GPCR led to stress fiber formation in NIH 3T3 cells. To examine the involvement of the Gα13-RhoA pathway in KSHV-GPCR-mediated NF-κB activation, HeLa cells were transfected with KSHV-GPCR alone and in combination with the regulator of G protein signaling (RGS) from p115RhoGEF or a dominant negative RhoA(T19N). Both constructs, as well as the C3 exoenzyme from Clostritium botulinum, partially reduced NF-κB activation by KSHV-GPCR, and by a constitutively active Gα13(Q226L). KSHV-GPCR-induced NF-κB activation is accompanied by increased secretion of IL-8, a function mimicked by the activated Gα13 but not by an activated Gα q(Q209L). These results suggest coupling of KSHV-GPCR to the Gα13-RhoA pathway in addition to other G proteins.

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