MOLECULAR EVOLUTION OF GPCRS: 26Rfa/GPR103

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   Figure 1

   Phylogenetic tree of the RFamide peptide family in vertebrates. Studies over the past decade have demonstrated that the brain of vertebrates produces a variety of RFamide peptides. To date, five groups have been identified within this family: the neuropeptide FF (NPFF) group, the prolactin-releasing peptide (PrRP) group, the gonadotropin-inhibitory hormone (GnIH) group, the kisspeptin group, and the 26RFa/QRFP group.
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   Figure 2

   Alignments of the amino acid sequences of identified 26RFa/QRFP peptides (A) and their precursor proteins (B) deduced from mammalian (human, bovine, rat, and mouse), avian (chicken, quail, and zebra finch), amphibian (Xenopus), and fish (goldfish) cDNAs. The predicted signal peptide sequences are underlined with a dashed line. <E represents pyroglutamic acid. The positions of identified mature peptides in the precursor proteins are underlined with solid lines. The human and Xenopus 26RFa/QRFP precursors may also generate a nine-amino acid peptide, termed 9RFa (boxed). Fully conserved amino acids are highlighted with red boxes and highly conserved amino acids with gray boxes respectively. The Lys (K)-Arg (R) dibasic processing sites in birds and Xenopus, the single Arg (R) putative processing sites in mammals and fish, and the Gly (G) C-terminal amidation signals are shown in bold. Gaps marked by hyphens were inserted to optimize homology. The GenBank accession numbers of these sequences are as follows: human 26RFa/QRFP, NP_937823; bovine 26RFa/QRFP, NP_937865; rat 26RFa/QRFP, NP_937843; mouse 26RFa/QRFP, NP_906269; chicken 26RFa/QRFP, XP_001235089; quail 26RFa/QRFP, BAI81890; zebra finch 26RFa/QRFP, BAK32798; Xenopus tropicalis 26RFa/QRFP, XP_002936227; and goldfish 26RFa/Qrfp, ACI46681.
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   Figure 3

   Synteny analysis around 26RFa/QRFP gene loci. Orthologous or paralogous genes are linked by horizontal lines. The 26RFa/QRFP genes are shown white in black boxes. The nucleotide position of each gene on the chromosome is shown under each gene. The GenBank accession numbers of 26RFa/QRFP genes are as follows: human 26RFa/QRFP, AB109625.1; mouse 26RFa/Qrfp, AB109628.1; chicken 26RFa/QRFP, XM_001235088.2; Xenopus tropicalis 26RFa/qrfp, XM_004916673.1; zebrafish 26RFa/qrfp, XP_00133883; medaka 26RFa/qrfp, XP_004073955.1.
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   Figure 4

   Demonstrated biological actions of 26RFa/QRFP–QRFPR system in vertebrates. Both 26RFa/QRFP and QRFPR have been found to exert a wide array of biological activities.
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   Figure 5

   Alignment of the amino acid sequences of the G protein-coupled receptor for 26RFa/QRFP, QRFPR, in mammals (human and rat), birds (chicken and zebra finch), frog (Xenopus), and fish (zebrafish). Fully conserved amino acids are highlighted with green boxes and highly conserved amino acids with gray boxes. Putative transmembrane domains (TMD) are underlined. The disulfide bridge between the two Cys (C) residues located in the first and second extracellular loops is indicated by a line. The Asp (D) residue in TMD2 involved in G protein coupling, the conserved Glu (E)-Arg (R) residues in the second intracellular loop, and the conserved Phe (F), Pro (P), and Asn (N) residues in TMD6 and TMD7 are represented by colored letters. A hyphen has been inserted to obtain optimal homology. The GenBank accession numbers of these sequences are as follows: human QRFPR, NP_937822; rat QRFPR, NP_937842; chicken QRFPR, NP_001120642; zebra finch QRFPR, NP_001243137; Xenopus tropicalis QRFPR, NP_001072295; and zebrafish Qrfpr, XP_001920042.
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   Figure 6

   Synteny analysis around QRFPR gene loci. Orthologous or paralogous genes are linked by horizontal lines. The QRFPR genes are shown white in black boxes. The nucleotide position of each gene on the chromosome is shown under each gene. The GenBank accession numbers of QRFPR genes are as follows: human QRFPR, JF810892.1; mouse Qrfpr1, BC096610.1; chicken QRFPR, NM_001127170.1; Xenopus tropicalis qrfpr, NM_001078827.1; and medaka qrfpr, XP_004080459.1. Ensembl genome database accession numbers are as follows: mouse Qrfpr2, ENSMUSG00000029917; zebrafish qrfpr1, ENSDARG00000039349; zebrafish qrfpr2, ENSDARG00000068422; zebrafish qrfpr3, ENSDARG00000092652; coelacanth qrfpr, ENSLACG00000016226; and sea lamprey qrfpr, ENSPMAG00000005451. GENSCAN (http://genes.mit.edu/GENSCAN.html) was used to predict putative coelacanth Qrfpr3 precursor protein.
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   Figure 7

   Phylogenetic analysis of QRFPR precursor proteins. Drosophila melanogaster peptide GPCR was used as an outgroup. NPY receptors are included in the phylogenetic tree as a reference group of vertebrate GPCR. Scale bar refers to a phylogenetic distance of 0.1 nucleotide substitutions per site. Numbers on the branches indicate bootstrap percentage following 1000 replications in constructing the tree. The GenBank accession numbers of the NPY1R genes are as follows: human NPY1R, NM_000909; mouse Npy1r, NM_010934; chicken NPY1R, NM_001031535; anole lizard NPY1R, XM_003221700; Xenopus laevisnpy1r, NM_001085879; zebrafish npy1r, NM_001102391; and Drosophila melanogaster peptide GPCR, AY217746.1.

• © 2014 Society for Endocrinology