Organization, 5′-flanking sequence and promoter activity of the rat GPC1 gene

• Published in: Gene Vol. 206; no. 2; pp. 255 - 261
• Main Authors: Asundi, Vinod K, Keister, Bonnie F, Carey, David J
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 12.01.1998
• Subjects: Animals; Base Sequence; Cells, Cultured; Cloning; Cloning, Molecular; DNA, Complementary - analysis; Exons; Expression; Fibroblasts; Genes; Heparan Sulfate Proteoglycans - chemistry; Heparan Sulfate Proteoglycans - genetics; Introns; Molecular Sequence Data; Promoter Regions, Genetic - genetics; Proteoglycan; Rats; Regulatory Sequences, Nucleic Acid; Schwann Cells; Transcription; Proteoglycan; oligo, oligodeoxyribonucleotide; Transcription; Cloning; kb, kilobase(s) or 1000 bp; bp, base pair(s); GPI, glycosylphosphatidylinositol; Expression; HSPG, heparan sulfate proteoglycan; bFGF, basic fibroblast growth factor; cDNA, DNA complementary to RNA; PCR, polymerase chain reaction; PDGF, platelet derived growth factor
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(97)00594-5

Glypicans are a member of a family of glycosylphosphatidylinositol anchored heparan sulfate proteoglycans that are expressed in cell and development specific patterns. Rat GPC1 cDNA probes were used to screen rat genomic libraries. Three overlapping genomic clones that contained the entire rat GPC1 gene were isolated. The rat GPC1 gene is approximately 15 kb in length and consists of eight exons interrupted by introns of varying lengths. Two of the introns are quite short, with lengths of 41 and 43 base pairs. Each exon–intron splice junction exhibited the consensus splice site sequence. Exon 1 encodes the putative signal peptide and the serine residue of the first putative heparan sulfate attachment site. The last exon encodes the cluster of three potential COOH-terminal heparan sulfate attachment sites, the putative GPI anchor and polypeptide cleavage site, and the 3′-untranslated region including the polyadenylation signal. One of the genomic clones extended approximately 2.8 kb 5′ of the exon 1 coding sequence, and is thus likely to contain sequences that regulate GPC1 gene expression. Sequence analysis of the 5′-flanking sequence revealed a lack of consensus TATA and CAAT boxes. A search for potential transcription factor binding sites revealed a number of such motifs, including Sp1 (GC box), NF- κB, and MyoD (E-box). This region of the rat GPC1 gene shows significant sequence homology to the 5′-flanking region of the human GPC3 gene. Functional promoter activity of the rat GPC1 sequence was demonstrated by its ability to drive the expression of a luciferase reporter gene in several cell types.