STRUCTURE, 5'-FLANKING SEQUENCE, AND CHROMOSOME LOCATION OF THE HUMANN-FORMYL PEPTIDE RECEPTOR GENE - A SINGLE-COPY GENE COMPRISED OF 2 EXONS ON CHROMOSOME 19Q.13.3 THAT YIELDS 2 DISTINCT TRANSCRIPTS BY ALTERNATIVE POLYADENYLATION
Dl. Haviland et al., STRUCTURE, 5'-FLANKING SEQUENCE, AND CHROMOSOME LOCATION OF THE HUMANN-FORMYL PEPTIDE RECEPTOR GENE - A SINGLE-COPY GENE COMPRISED OF 2 EXONS ON CHROMOSOME 19Q.13.3 THAT YIELDS 2 DISTINCT TRANSCRIPTS BY ALTERNATIVE POLYADENYLATION, Biochemistry, 32(16), 1993, pp. 4168-4174
The N-formyl peptide chemoattractant receptor (fMLF-R) is a cell-surfa
ce, G-protein-coupled glycoprotein that mediates the directed locomoti
on of neutrophils upon binding N-formylated peptides. The fMLF-R is en
coded primarily by a 1.6-kb mRNA in differentiated HL-60 and U937 cell
s, although larger less abundant transcripts are present. To study the
origin of different fMLF-R transcripts, the genetic linkage of chemot
actic receptor genes, and the regulation of fMLF-R gene expression, we
determined the copy number, chromosomal location, structural organiza
tion, and 5'-flanking sequence of the human fMLF-R gene. BamHI restric
tion fragments derived from a human fMLF-R genomic cosmid clone were i
solated, subcloned, and sequenced. These data indicate that the fMLF-R
structural gene is approximately 7.5 kb in length and is comprised of
two exons separated by an approximately 5.0-kb intron. The first exon
encodes 66 bp of the 5'-untranslated sequence, while exon 2 encodes t
he coding and 3'-untranslated sequences. The genomic organization of t
he fMLF-R gene is similar to that of the adrenergic beta-1 and beta-2
G-protein-coupled receptor genes in that the coding sequence is contai
ned in a single exon. The different 3'-untranslated sequences observed
in fMLF-R cDNA clones are contiguous in the genomic structure, thereb
y indicating that these clones are derived in part by alternative poly
adenylation. 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 dbc
AMP-differentiated U937 RNA indicated a single transcriptional initiat
ion site. Sequence analysis 5' of the transcriptional initiation site
indicated possible cis-acting motifs that may regulate fMLF-R gene exp
ression. These included AP-1 and CK-2 consensus sequences that bind nu
clear factors of the Fos/Jun family and NF-GMb, respectively.