Identification and analysis of the gene encoding human PC2, a prohormone convertase expressed in neuroendocrine tissues.
In recent studies we have identified PC2 and PC3, members of a family of serine proteases that are related structurally to subtilisin, and have provided evidence that these are involved in the tissue-specific processing of prohormones and neuropeptides. PC2 is expressed at high levels in the islets of Langerhans, where it participates in the processing of proinsulin to insulin (S.P.S. and D.F.S., unpublished data). To evaluate the regulated expression of the human PC2 (hPC2) gene we have analyzed its structure and characterized its promoter. A map of the gene was constructed by using 11 clones isolated from two human genomic DNA libraries. The gene spans greater than 130 kilobase pairs and consists of 12 exons. Comparison with the structure of the gene encoding human furin, another member of this superfamily, revealed a high degree of conservation of exon-intron junctions. The hPC2 gene was localized to chromosome 20, band p11.2. The 5' flanking region of the hPC2 gene is very G+C-rich and contains six potential Sp1 binding sites but no TATA or CAAT box. Expression of chloramphenicol acetyltransferase reporter fusions containing the putative promoter region was observed to occur in beta TC-3 mouse insulinoma cells but not in HepG2 human hepatoma cells, consistent with the known tissue-specific pattern of expression of the hPC2 gene. Analysis of the level of chloramphenicol acetyltransferase activity with several deletion mutants identified the region from -1100 to -539 from the translation start site as essential for hPC2 promoter activity.
Ohagi
S
S
Howard Hughes Medical Institute, Department of Biochemistry and Molecular Biology, Chicago, IL 60637.
LaMendola
J
J
LeBeau
M M
MM
Espinosa
R
R
Takeda
J
J
Smeekens
S P
SP
Chan
S J
SJ
Steiner
D F
DF
D000595:Amino Acid Sequence; D001483:Base Sequence; D002890:Chromosomes, Human, Pair 20; D003001:Cloning, Molecular; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D045706:Proprotein Convertase 2; D011498:Protein Precursors / Q000378:metabolism; D015183:Restriction Mapping; D016415:Sequence Alignment; D012697:Serine Endopeptidases / Q000235:genetics*
D011498:Protein Precursors; D012697:Serine Endopeptidases; D045706:Proprotein Convertase 2