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• W2318007048 abstract "41 Background: Kaposi's sarcoma-associated herpesvirus (KSHV/HHV 8) encodes a GPCR which is most homologous to cellular chemokine receptors, in particular to IL-8 (CXCR1 and CXCR2). Expression of the viral GPCR gene has been shown in PELs and in KS by RT-PCR analysis. This receptor is constitutively active, signaling in the absence of ligand. Furthermore, KSHV GPCR stimulates cellular proliferation, is transforming in NIH3T3 cells, and induces VEGF-mediated angiogenesis. Sequencing and functional characterization in our laboratory has been performed using only one clone obtained from a PEL. A second reported sequence of this ORF diverged significantly in the fourth transmembrane domain from our sequence (Guo et al., Virology 228, 1997). Thus, our first aim was to sequence and determine the function of the KSHV GPCR cloned from other KS and PEL specimens to assess whether the constitutive activation and other biologic features were conserved. Our second aim was to characterize the pattern of expression and the transcripts encoding this gene. Methods: KSHV GPCR was cloned from two PEL specimens, one PEL cell line (BC-3), and two KS specimens, followed by sequence analysis. Signaling was measured by inositol phosphate accumulation. The KSHV-GPCR cDNA was cloned from a TPA-treated BC-1 cDNA library. The pattern of expression was characterized in BC-1, BC-2 and BC-3 cell lines by Northern blot analysis before and after TPA induction, and/or treatment with foscarnet or cycloheximide. Results: The DNA sequence of the KSHV-GPCR gene obtained from five independent sources (two PEL and two KS specimens, and one PEL cell line) showed two or three base differences when compared to the prototype sequence, but none of these changes led to alterations at the amino acid sequence level. As expected, all clones were similar to the prototype in their signaling characteristics, and all were constitutively active. The cDNA clones obtained contained a 2.6 kb inserf, corresponding to the size identified by Northern blotting. This is a bicistronic message, containing the K14 ORF upstream from the GPCR. A Kozak consensus sequence for initiation of translation is present in the GCPR, but not in the K14 ORF, suggesting that the GPCR can be translated in spite of the upstream ORF present in the mRNA. Northern blot analysis in the PEL cell lines showed that the KSHV GPCR is an immediate early lytic gene that continues to be transcribed at an increased level throughout the lytic cycle. A low level of KSHV GPCR mRNA is seen in mainly latently infected PEL cell lines, as well as in PEL and KS specimens. Conclusions: The KSHV GPCR gene is transcribed as a bicistronic message with an immediate early lytic pattern. The KSHV GPCR protein sequence is completely conserved among different KS and PEL tissues, demonstrating that the constitutive activity, as well as the transforming and angiogenic properties of this viral receptor are not particular to the clone originally examined. The high degree of conservation suggests that this protein has an important function in the life cycle of KSHV and in the pathogenesis of KS and PEL." @default.
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• W2318007048 title "CHARACTERIZATION OF KSHV/HHV-8 G PROTEIN-COUPLED RECEPTOR (GPCR) TRANSCRIPTS: STRUCTURAL AND FUNCTIONAL CONSERVATION AMONG KAPOSI'S SARCOMA AND PRIMARY EFFUSION LYMPHOMA." @default.
• W2318007048 doi "https://doi.org/10.1097/00042560-199804010-00056" @default.
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