Roles of GP33, a guinea pig cytomegalovirus-encoded G protein-coupled receptor homolog, in cellular signaling, viral growth and inflammation in vitro and in vivo
Fig 1
cDNA structure and expression of GP33.
(A) The GP33 cDNA structure determined by RACE analyses is depicted with a TATA box and a poly A signal (pA). The numbers in the GPCMV genome positions are based on Kanai et al. [33]. The ORF predicted initially as GP33 (hatched box; GP33s) and the ORF determined by RACE (closed boxes; GP33) are shown. An open arrowhead indicates the location of the DRY motif. “ins TGAT” is the mutation containing a stop codon and a 1-bp insertion to generate the GP33-defective virus, Δ33. (B) Multiple sequence alignment of GP33 homologs, HCMV UL33, MCMV M33, and RCMV R33, was performed by ClustalW analysis (DDBJ, ver. 2.1) with the default parameters except for GAP OPEN 100, and the obtained bootstrapped tree is depicted by TreeViewX software. (C) Localization of GP33-EGFP fusion protein in COS-7 cells transiently transfected with pEGFP-GP33. (D) Lysates of cells transfected with pcDNA3 empty vector (lane 1) or pcDNA-GP33F (lane 2) were separated on SDS-10% polyacrylamide gel. Proteins in the gel were transferred to polyvinylidene difluoride membrane (Millipore), and reacted with monoclonal anti-FLAG tag antibody followed by peroxidase-conjugated anti-mouse IgG. An arrowhead indicates the expected full-length GP33 band.