Structure, 5′-Flanking Sequence, and Chromosome Location of the Human N-Formyl Peptide Receptor Gene. A Single-Copy Gene Comprised of Two Exons on Chromosome 19q.l3.3 That Yields Two Distinct Transcripts by Alternative Polyadenylation

David L. Haviland, Angela C. Borel, Daniel T. Fleischer, Joie C. Haviland, Rick A. Wetsel

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

The N-formyl peptide chemoattractant receptor (fMLF-R) is a cell-surface, G-protein-coupled glycoprotein that mediates the directed locomotion of neutrophils upon binding N-formylated peptides. The fMLF-R is encoded primarily by a 1.6-kb mRNA in differentiated HL-60 and U937 cells, although larger less abundant transcripts are present. To study the origin of different fMLF-R transcripts, the genetic linkage of chemotactic receptor genes, and the regulation of fMLF-R gene expression, we determined the copy number, chromosomal location, structural organization, and 5′-flanking sequence of the human fMLF-R gene. BamHI restriction fragments derived from a human fMLF-R genomic cosmid clone were isolated, subcloned, and sequenced. These data indicate that the fMLF-R structural gene is ~7.5 kb in length and is comprised of two exons separated by an ~5.0-kb intron. The first exon encodes 66 bp of the 5′-untranslated sequence, while exon 2 encodes the coding and 3′-untranslated sequences. The genomic organization of the fMLF-R gene is similar to that of the adrenergic β-1 and β-2 G-protein-coupled receptor genes in that the coding sequence is contained in a single exon. The different 3′-untranslated sequences observed in fMLF-R cDNA clones are contiguous in the genomic structure, thereby indicating that these clones are derived in part by alternative polyadenylation. Southern blot analysis using human X hamster somatic cell hybrids and in situ hybridization indicated that the h-fMLF-R gene is located on chromosome 19q13.3. Primer extension experiments using dbcAMP-differentiated U937 RNA indicated a single transcriptional initiation site. Sequence analysis 5′ of the transcriptional initiation site indicated possible cis-acting motifs that may regulate fMLF-R gene expression. These included AP-1 and CK-2 consensus sequences that bind nuclear factors of the Fos/Jun family and NF-GMb, respectively.

Original languageEnglish (US)
Pages (from-to)4168-4174
Number of pages7
JournalBiochemistry
Volume32
Issue number16
DOIs
StatePublished - Apr 1 1993

ASJC Scopus subject areas

  • Biochemistry

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