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J. Biol. Chem., Vol. 276, Issue 49, 45979-45987, December 7, 2001
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B and Secretion of Interleukin-8
Induced by the G Protein-coupled Receptor of Kaposi's
Sarcoma-associated Herpesvirus Involve G
13 and RhoA*
,
From the Department of Pharmacology, College of Medicine,
University of Illinois, Chicago, Illinois 60612
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 
B (NF-B) and secretion of interleukin-8, and this
response involves activation of G
13 and RhoA. The
induced expression of a NF-B luciferase reporter was partially
reduced by pertussis toxin and the G
scavenger transducin, and
enhanced by co-expression of G13 and to a lesser extent,
Gq. These results indicate coupling of KSHV-GPCR to
multiple G proteins for NF-B activation. Expression of KSHV-GPCR led
to stress fiber formation in NIH 3T3 cells. To examine the involvement
of the G13-RhoA pathway in KSHV-GPCR-mediated NF-B
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-B activation by KSHV-GPCR, and by a
constitutively active G13(Q226L). KSHV-GPCR-induced NF-B activation is accompanied by increased secretion of IL-8, a
function mimicked by the activated G13 but not by
an activated Gq(Q209L). These results suggest
coupling of KSHV-GPCR to the G13-RhoA pathway in
addition to other G proteins.
Present address: Dept. of Microbiology and Immunology, University
of Iowa.
§
Recipient of a Biomedical Science Grant from the Arthritis Foundation.
¶
To whom correspondence should be addressed. Tel.:
312-996-5087; E-mail: yer@uic.edu.
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