Molecular Organization of the Human Cathepsin D Gene

Abstract

A 16-kb fragment of human DNA containing the cathepsin D (CATD) gene was isolated. Nucleotide sequencing, primer extension, protection from mung bean nuclease, and promoter activity assays were used to characterize the gene. The transcribed portion of the gene is about 11,000 bp and is organized into 9 exons analogous with the human pepsinogen A gene. Human pepsinogen A and CATD proteins have 42% sequence identity, while the two cDNAs are 55.7% identical. The positions of the splice junctions are fully conserved in these two genes. The noncoding sequences of the two genes are dissimilar. We report the nucleotide sequence of an Eco RI-Bam HI fragment that contains the transcription initiation site. The promoter region contains no TATA and CCAAT boxes, but five potential Sp1 binding sites (one of them in the first intron) and four AP-2 binding sites (two of them in the first intron). In COS-1 cells, the region containing the three proximal Sp1 sites possesses the bulk of the promoter activity of the 5'-flanking sequence. The transcription start site of the CATD gene is localized within a CpG cluster. In the interval -390 through +450, the content of CpG is 5.8 times above the average throughout the human genome.

Rice Aspartic Proteinase, Oryzasin, Expressed During Seed Ripening and Germination, has a Gene Organization Distinct from Those of Animal and Microbial Aspartic Proteinases

“…An intron exists in the 5'-noncoding region of the oryzasin 1 gene. This has never before been seen in AP genes [18][19][20][21][22][23][24]371. As ; RR, rat renin [20] ; BC, bovine chymosin [21]; YPA, yeast proteinase A [37]; RNAP, [23]; PEPA, aspergillopepsin A [24].…”

“…4). Animal AP genes, such as human cathepsin D [19], rat renin [20], bovine chymosin [21] (Fig. 4) and human pepsinogen genes [18], are all made up of nine exons and eight introns, with the only exception being that of the human renin gene which has 10 exons and nine introns [22].…”

Human cathepsin D.

“…It consists of 9 exons and contains 11.106 base pairs. For a complete sequence of cathepsin D gene please refer to paper by Roedecker et al [8]. Cathepsin D gene transcription leads to the formation of mRNA containing 1.988 bases.…”

Molecular Mechanism of Inhibition of Estrogen-Induced Cathepsin D Gene Expression by 2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) in MCF-7 Cells

“…Levels of acetylated product were quantitated by a Betagen Betascope 603 blot analyzer imaging system and visualized by autoradiography. Secreted alkaline phosphatase activity was determined essentially as described elsewhere (5,63). One milliliter of medium was obtained at the appropriate times and incubated for 10 min at 65ЊC.…”

“…The human estrogen receptor (hER) expression plasmid was provided by Ming-Jer Tsai (Baylor College of Medicine, Houston, Tex.). The pB12/pL/pac (Seap-pac) plasmid containing the cathepsin D promoter (Ϫ296/ϩ57) fused to an alkaline phosphatase gene was used to study the promoter activity (63). The pBC12/S1/pac (S1) plasmid containing no promoter and the pBC12/RSV/pac (RSV) plasmid containing a Rous sarcoma virus promoter fused to the alkaline phosphatase gene were used as negative and positive controls, respectively.…”