Human renin gene: structure and sequence analysis.

Abstract

The complete protein precursor of human kidney renin has been determined from the sequence of cloned genomic DNA. The gene spans 12 kilobases of DNA and is interrupted by eight intervening sequences. The nine regions (exons) encoding the protein were mapped with a mouse renin cDNA probe, synthetic oligonucleotide probes, and by hybridization of genomic restriction fragments to a 1600-nucleotide human kidney mRNA. The predicted 403-amino acid preprorenin consists of mature renin and a 66-residue amino-terminal prepropeptide. The DNA sequence 5' to the first exon indicates the location of a transcriptional promoter (T-A-T-A-A-A) for a mRNA encoding preprorenin. An additional transcriptional promoter site is located within the first intron, which, if used, would express a shortened nonsecreted prorenin. The structure of the human renin gene is similar to that of human pepsinogen, a closely related aspartyl protease enzyme. This observation suggests that renin and pepsinogen have a common evolutionary origin.Renin is an endocrine hormone catalyzing the first step in a cascade of factors that modulate arteriole blood pressure. It hydrolyzes a single peptide bond in the circulating globulin, angiotensinogen, releasing the amino-terminal decapeptide angiotensin I. The absolute specificity of renin is in contrast to other aspartyl proteases, which act on a broad range of substrates (1). The characterization of this specificity is of considerable pharmaceutical and medical interest. However, detailed biochemical analysis of the protein has been limited because the hormone is produced in such small amounts by its known physiological source, the juxtaglomerular cells of the kidney cortex. Renin purified from human tissue has shown variations in molecular weight and amino acid composition (2-4).We report here the isolation and sequence analysis of the human renin gene.EXPERIMENTAL PROCEDURES Plaque Screening. A library of bacteriophage Charon 4A containing human fetal DNA (5) was grown in Escherichia coli strain LE392 (6). Nitrocellulose filter (Schleicher & Schuell) replicas of -300,000 plaques were incubated in 2x NaCl/Cit (lx NaCl/Cit is 0.15 M NaCl/0.015 M Na citrate)/0.1% NaDodSO4/5x Denhardt's solution (lx Denhardt's solution is 0.02% polyvinylpyrrolidone/Ficoll (Pharmacia)/bovine serum albumin) at 55°C overnight and then hybridized (106 cpm per filter) to a mouse renin probe [labeled to greater than 108 cpm/,g of DNA by nick-translation (7) in the presence of [a-32P]dATP and dCTP (New England Nuclear)] at 55°C for 24 hr in 2x NaCl/Cit/0.1% NaDodSO4/1x Denhardt's solution/10% dextran sulfate (Pharmacia). Filters were washed at 50°C in 0.1x NaCl/Cit/0.1% NaDodSO4 prior to autoradiography. Positive plaques were purified by standard methods (8). Cloning procedures were according to National Institutes of Health guidelines.Hybridization to Filter-Bound mRNA. RNA was purified (9) from human kidneys (obtained immediately postmortem or from surgical nephrectomy) and the poly(A)+ mRNA was fractionated using poly(U)-Se...

Loss of heterozygosity for the short arm of chromosome 1 in human neuroblastomas: correlation with N-myc amplification.

“…We used a panel of 19 chromosome-i-specific DNA probes (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22), and the order of these probes was determined by genetic linkage in the Centre d'Etude du Polymorphisme Humain reference panel families (10). Of these probes, eight define known gene loci-proatriodilatin (PND) (11), a-fucosidase (FUCAl) (12), the protooncogene c-fgr (FGR) (13), the protooncogene L-myc (MYCL) (14), a-amylase (AMY1) (15), ,3 nerve growth factor (NGFB) (16), anti-thrombin III (AT3) (17), and renin (REN) (18); two probes define chromosome-i-specific repetitive sequences-D1Z2 (19) and D1S57 (20); seven probes were isolated from a flow-sorted chromosome 1 library (D1S15, D1S16, D1S17, D1S18, D1S19, D1S21, and D1S22) (10), and two probes define anonymous DNA sequences-CRI-L336 (D1S47) (21) and D1S2 (22). A locus is considered informative for a particular patient when the constitutional DNA from that patient displays two different alleles (i.e., heterozygosity at that locus).…”

Diabetic complications: A role for the prorenin–(pro)renin receptor–TGF-β1 axis?

“…With the cloning of the renin gene in 1984, prorenin was proven to be the precursor of renin (Hobart et al, 1984). A 43-amino acid N-terminal propeptide explains the absence of enzymatic activity of prorenin as compared to renin.…”

Evidence for a local renin angiotensin system in primary cultured human preadipocytes

“…20,21 Renin primers were 5 H -TACAACAGGAATTCCCAATC-3 H (exon 5a6) and 5 H -GACACC-CCCTTCATTTGAAT-3 H (exon 6a7) so as to amplify bases 736 to 859 of the human renin cDNA. 22 RBP primers were 5 H -CAG-CGCGACAGGA-CATGGAGAAA-3 H (exon 1a2) and 5 H -CAATACATC-CATAC-CTGCCTCCC-3 H (exon 3a4) so as to amplify bases 17 to 227 of the human RBP cDNA. 23,24 ACE primers were 5 H -AGGAGCTGGAGGAGTACAAC-3 H (exon 16a17) and 5 H -CCCTG-CATCTACATAGC-CATT-GA-3 H (exon 17a18) so as to amplify bases 2225 to 2488 of the human ACE cDNA.…”