The Structure of the Prostaglandin EP4 Receptor Gene and Related Pseudogenes

The EP4 prostaglandin receptor (EP4R) is a member of the seven transmembrane receptor superfamily. We have obtained the human EP4 receptor gene sequence and determined its structure relative to EP4R cDNA synthesized from peripheral blood lymphocytes. The EP4R gene spans approximately 22 kb and consi...

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Bibliographic Details
Published in:Genomics (San Diego, Calif.) Calif.), 1996-07, Vol.35 (1), p.182-188
Main Authors: Foord, Steven M., Marks, Bruno, Stolz, Martin, Bufflier, Emmanuel, Fraser, Neil J., Lee, Melanie G.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
Biological and medical sciences
Cloning, Molecular
DNA, Complementary - genetics
Exons - genetics
Fundamental and applied biological sciences. Psychology
Gene Library
Genes
Genes. Genome
Humans
Introns - genetics
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Polymerase Chain Reaction
Pseudogenes
Receptors, Prostaglandin E - genetics
Receptors, Prostaglandin E, EP4 Subtype
Sequence Alignment
Sequence Homology, Nucleic Acid
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Summary:The EP4 prostaglandin receptor (EP4R) is a member of the seven transmembrane receptor superfamily. We have obtained the human EP4 receptor gene sequence and determined its structure relative to EP4R cDNA synthesized from peripheral blood lymphocytes. The EP4R gene spans approximately 22 kb and consists of three exons separated by two introns. The first exon (530 bp) is noncoding. After an intron of 472 bp, the second exon contains a short (43 bp) 5′ sequence before a 289-amino-acid open reading frame (ORF). An 11.5-kb intron is found at the end of transmembrane 6, and the rest of the ORF is in exon 3. The gene structure is analogous to those of the thromboxane, PGI, and PGD receptors. The deduced initiation site does not contain a conventional TATA box but is 70% GC-rich and contains CCAAT boxes, SP1 and AP2 motifs, and motifs consistent with activation by proinflammatory cytokines. Southern blot analysis of human genomic DNA shows two genes with homology to the EP4R gene. Both appear to be pseudogenes with 70% amino acid identity to the EP4R up to the “ERY” sequence at the end of transmembrane 3, where anAlu-like repetitive sequence element was found. The ORF sequence is also interrupted by a stop codon. The pseudogenes differ in that one contains a second “repetitive element” (a line 1 repeat) in the 5′ end of the ORF. Northern blot analysis of human mRNA using a pseudogene probe showed hybridization only to the EP4 receptor transcript. PCR also failed to detect expression of either pseudogene. This study defines the gene structure of EP4R and suggests the existence of two related pseudogenes.
ISSN:0888-7543
1089-8646
DOI:10.1006/geno.1996.0337