Structural characterization of the human carboxypeptidase D gene and its promoter.
Human carboxypeptidase D (CPD) is a 180-kDa type I membrane protein with three tandem active site domains. CPD is a B-type (or kininase I-type) carboxypeptidase that cleaves C-terminal basic residues from proteins and peptides, such as Arg9 from bradykinin. The human carboxypeptidase D (CPD) gene was found to encompass approximately 88.3 kb of genomic sequence, containing 21 exons ranging in size from 65 to 1813 bp, and 21 introns ranging in size from 112 bp to 35.6 kb. Although CPD and CPM belong to the same metallocarboxypeptidase subfamily, their intron/exon structures differ significantly. Multiple transcription start sites were found in the CPD gene within a GC-rich sequence lacking the typical TATA box, but containing three GC boxes. Luciferase reporter assays with various size constructs containing the promoter region upstream of the start sites showed that it was active in three different cell lines, especially in the human hepatoma cell line HepG2 and the human monocytic cell line THP-1, which have high constitutive expression of CPD.
Timblin
Barbara
B
Department of Pharmacology, College of Medicine, University of Illinois, Chicago, IL 60612, USA.
Rehli
Michael
M
Skidgel
Randal A
RA
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002268:Carboxypeptidases / Q000235:genetics*; D002460:Cell Line; D006224:Cricetinae; D005091:Exons / Q000235:genetics; D005796:Genes / Q000235:genetics*; D006801:Humans; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000235:genetics*
D002268:Carboxypeptidases; C078874:carboxypeptidase D