Cloning and characterization of the guinea pig neutrophil cationic peptide-1 and -2 genes.
Guinea pig neutrophils contain the antimicrobial cationic peptides GNCP-1 and GNCP-2 in the granules. Using cDNA probes, four different GNCP gene clones were isolated from a guinea pig genomic library. Two clones encoded GNCP-1 and other two clones encoded GNCP-2. The nucleotide sequence of GNCP-1 and GNCP-2 gene clones were highly homologous (> 97%) in the region sequenced. The GNCP-1 and GNCP-2 genes spanned 3 kb, and consisted of three exons and two introns. Exon 1 encoded the 5' untranslated region, exon 2 encoded the prepro-peptide region, and exon 3 encoded the mature peptide region and the 3' untranslated region. The transcription start site was located to a thymidine residue 93 bp upstream of the start codon (ATG) of GNCP-1 or GNCP-2 mRNA, and TATA-like sequence was located 24 to 30 bp upstream of the transcription start site. Glucocorticoid regulating element-like sequence was found in the 5' flanking region of the GNCP-1 or GNCP-2 gene, and the GNCP gene transcription rate of bone marrow cells was upregulated by dexamethasone treatment. Together these observations indicate that GNCP-1 and GNCP-2 are encoded by the very homologous but different genes, and that GNCP gene expression is regulated by glucocorticoid.
Nagaoka
I
I
Department of Biochemistry, Juntendo University, School of Medicine, Tokyo, Japan.
Nonoguchi
A
A
Yamashita
T
T
D000818:Animals; D023181:Antimicrobial Cationic Peptides; D001483:Base Sequence; D001798:Blood Proteins / Q000096:biosynthesis / Q000235:genetics*; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003907:Dexamethasone / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D006168:Guinea Pigs / Q000235:genetics*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; C030509:neutrophil basic proteins; D003907:Dexamethasone