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F Soubrier

Publications and source records attributed to F Soubrier.

At least 73 records · Page 4Linked to original sources
PubMed1995

Renin gene expression in the aging kidney: effect of sodium restriction.

The activity of the renin-angiotensin system as well as the ability of the kidney to retain sodium following salt restriction are reduced with age. The relationship between these age-related changes in renal function and the renin gene expression was presently investigated. The concentrations of renin and its mRNA were measured in kidney of 10- and 30-month-old control female WAG/Rij rats and of animals which were salt restricted for 4 days. In the senescent rats, the kidney renin concentration, like the plasma concentration of angiotensin II, was half that in adult rats. The intrarenal content of renin mRNA did not differ between 10- and 30-month-old animals, suggesting that the transcriptional rate of the renin gene is unchanged with age. During the early phase of adaptation to sodium depletion, the systemic angiotensin II concentration was not modified in either age groups. Four-days salt restriction did not significantly change the renal storage of renin. In contrast, this short term salt restriction induced a 2.3-fold increase in the renin mRNA in adult kidney, and a 1.9-fold increase in the senescent kidney. These data suggest that the age-related decrease in renal concentration of renin is linked to a modification in the rate of translation of renin mRNA, or to an alteration in the protein maturation. The difference in adaptation to the early phase of salt restriction with age should not be linked to changes in renin gene transcription, but more likely to a change in the tissue response to the local renin-angiotensin system.

Adaptation, Physiological↗
PubMed1995

Human mineralocorticoid receptor genomic structure and identification of expressed isoforms.

Most of the known effects of aldosterone are mediated by the mineralocorticoid receptor, an intracellular receptor belonging to the steroid/thyroid hormone/retinoic acid receptor superfamily. We determined the genomic structure of the human MR (hMR) and identified 10 exons in the gene, including two exons (1 alpha and 1 beta) that encode different 5'-untranslated sequence. Expression of the two different hMR variants is under the control of two different promoters that contain no obvious TATA element, but multiple GC boxes. Our results indicate that hMR expression is regulated by alternative promoters perhaps in a tissue- or developmental-specific manner.

Bacteriophage M13↗
PubMed1995

Aliphatic nitrilase from a soil-isolated Comamonas testosteroni sp.: gene cloning and overexpression, purification and primary structure.

An aliphatic nitrilase, active on adiponitrile and cyanovaleric acid, was identified and purified from Comamonas testosteroni sp. (Ct). Oligodeoxyribonucleotide probes were designed from limited amino acid (aa) sequence information and used to clone the corresponding gene, named nitA. High homologies were found at the aa level between Ct nitrilase and the sequences of known nitrilases. Multi-alignment of sequenced nitrilases suggests that Cys163 of Ct plays an essential role in the active site. This hypothesis is strengthened by molecular studies on nitrilases from Alcaligenes faecalis JM3, and Rhodococcus rhodochrous J1 and K22 [Kobayashi et al., Proc. Natl. Acad. Sci. USA 90 (1993) 247-251; J. Biol. Chem. 267 (1992) 20746-20751; Biochemistry 31 (1992) 9000-9007]. Large amounts of an active recombinant enzyme could be produced in Escherichia coli when nitA was overexpressed together with the E. coli groESL genes.

Amino Acid Sequence↗
PubMed1995

Cloning and expression of an evolutionary conserved single-domain angiotensin converting enzyme from Drosophila melanogaster.

Mammalian somatic angiotensin converting enzyme (EC 3.4.15.1, ACE) consists of two highly homologous (N- and C-) domains encoded by a duplicated gene. We have identified an apparent single-domain (67 kDa) insect angiotensin converting enzyme (AnCE) in embryos of Drosophila melanogaster which converts angiotensin I to angiotensin II (Km, 365 microM), removes Phe-Arg from the C terminus of bradykinin (Km, 22 microM), and is inhibited by ACE inhibitors, captopril (IC50 = 1.1 x 10(-9) M) and trandolaprilat (IC50 = 1.6 x 10(-8) M). We also report the cloning and expression of a Drosophila AnCE cDNA which codes for a single-domain 615-amino acid protein with a predicted 17-amino acid signal peptide and regions with high levels of homology to both the N- and C-domains of mammalian somatic ACE, especially around the active site consensus sequence. Northern analysis identified a single 2.1-kilobase mRNA in Drosophila embryos, and Southern analysis of Drosophila genomic DNA indicates that the insect gene is not duplicated. When expressed in COS-7 cells, the AnCE protein is a secreted enzyme, which converts angiotensin I to angiotensin II and is inhibited by captopril (IC50 = 5.6 x 10(-9) M) and trandolaprilat (IC50 = 2 x 10(-8) M). The evolutionary significance of these results is discussed.

Amino Acid Sequence↗
PubMed1995

Lack of evidence for linkage of the endothelial cell nitric oxide synthase gene to essential hypertension.

BACKGROUND: The basal release of nitric oxide by the endothelium plays an important role in regulating blood flow and pressure and mediates most of the endothelium-dependent vasodilation. Impairment of nitric oxide production by specific inhibitors increases blood pressure in humans, and several reports suggest that hypertensive subjects have a blunted endothelium-dependent vasodilatation that might be secondary to decreased nitric oxide production from the vessel wall. METHODS AND RESULTS: To determine whether the endothelial nitric oxide synthase gene is involved in human essential hypertension, we identified informative biallelic and multiallelic markers of this locus and performed case-control and linkage studies in hypertensive subjects and normotensive control subjects. We used the affected sib pair method to test for potential linkage in 145 hypertensive pedigrees (269 sib pairs, 346 subjects) with a highly polymorphic marker of the nitric oxide synthase gene (polymorphism information content of 92%). There was no evidence for linkage among affected siblings. The 95% upper confidence limit of this value suggests that at most 1% of alleles in excess of expected are shared. We also identified two informative biallelic markers of this gene to perform a case-control study on white hypertensive and normotensive subjects. Similar genotype distributions between the two groups were noted for both markers. Estimated haplotype frequencies by maximum likelihood methods combining the two biallelic markers were also similar in both groups. CONCLUSIONS: These findings do not suggest that common molecular variants of the endothelial nitric oxide synthase gene are involved in essential hypertension.

Adult↗
PubMed1995

Molecular characterization of the mineralocorticoid receptor in pseudohypoaldosteronism.

Pseudohypoaldosteronism (PHA) is characterized by salt-wasting and failure to thrive in the newborn, accompanied by high urinary levels of sodium despite hyponatremia, hyperkalemia and metabolic acidosis, elevation of plasma renin activity, and high plasma aldosterone levels. PHA patients are resistant to mineralocorticoid administration, but their symptoms ameliorate after a period of sodium supplementation, which can be discontinued in older subjects. Binding studies performed on mononuclear leukocytes of the family members affected by the disease have shown the absence of binding of [3H]aldosterone to the mineralocorticoid receptor (MR) in mononuclear leukocytes in two siblings and a marked reduction in another sibling and the father, suggesting either the absence of MR or a defect in the ligand binding domain of the MR in these patients. Molecular analysis of the MR in the members of this family did not reveal any major rearrangement or deletion of the MR gene. In addition, no mutation was found in the entire MR coding sequence by RT-PCR and direct sequencing of MR mRNA, and the semiquantitative RT-PCR analysis of the MR mRNA of one affected patient failed to show any quantitative abnormality in MR expression. These results do not exclude a molecular abnormality present in the MR gene being responsible for PHA. However, they indicate that in this family PHA is not related to a modification of the MR primary structure or to a major abnormality in MR expression.

DNA, Complementary↗
PubMed1995

A genetic study of angiotensin I-converting enzyme levels in human semen.

The plasma level of angiotensin I-converting enzyme (ACE) has been shown to be under genetic control. An insertion/deletion polymorphism in the ACE gene is associated with differences in the level of ACE in the plasma and inside T-lymphocytes. An ACE isoform is present in large amounts in spermatozoa and is expressed under an alternative, germ cell-specific promoter, whereas ACE present in the seminal fluid is the somatic form of ACE. We have investigated the effect associated with the I/D polymorphism on the level of ACE in seminal fluid and in spermatozoa. No differences in the level of ACE measured in the seminal fluid or in the spermatozoa were associated with the ACE I/D genotypes. We conclude that the modulation of expression associated with the I/D polymorphism is restricted to the somatic ACE promoter. These results also suggest that if one allele modulating the expression of ACE was under positive selection pressure, it was not through an effect on the semen concentration of ACE.

Alleles↗
PubMed1995

Recent advances in knowledge of the structure and function of the angiotensin I converting enzyme.

AIM: To review the structure and function of the angiotensin I converting enzyme (ACE), focusing on recent results from studies using a wide range of molecular biological techniques. ACE STRUCTURE AND FUNCTION: ACE is an ectoenzyme expressed as two isoenzymes in mammals, a larger somatic form found in endothelial, epithelial and neuronal tissues and a smaller form in germinal tissues. Both forms have similar enzymatic activities but differ in size and immunological properties. The somatic form of ACE is composed of two highly homologous domains (amino and carboxyl domains) while the germinal form contains only one domain. Somatic ACE has two functional catalytic sites, both dependent on a zinc cofactor. Each ACE domain has also been shown to interact differently with competitive inhibitors. MECHANISM OF ACE ANCHORAGE AND SOLUBILIZATION: The mechanism for anchoring ACE to the cell membrane has also been reported, and the solubilization step outlined. The relationship between the membrane-bound and soluble forms has been investigated, and the physiological relevance of this mechanism discussed. GENETIC STRUCTURE: The structure of the ACE gene has been determined and the distribution in cells and different tissues has been reported in various studies. CONCLUSION: All results have indicated that there are important functional and structural differences between the two domains, but at present ACE cannot be considered a true bifunctional enzyme, even though an exclusive substrate has been identified for the amino domain.

Animals↗
PubMed1995

The genetic basis of hypertension.

Genetic approaches are setting the stage for a new understanding of the biological mechanisms underlying blood pressure regulation and hypertension. Recently, substantial progress has been made in elucidating the molecular mechanisms that cause several rare forms of hypertension, including hypertension arising from mutations in the 11 beta-hydroxylase gene, Liddle's syndrome, and glucocorticoid-suppressible aldosteronism. New results have been obtained on the possible role of the angiotensinogen gene, and other candidate genes, such as the angiotensin II receptor Type I gene and the Sa gene, in human hypertension. Investigation of experimental models of hereditary hypertension have also been important in unravelling the genetic complexity of the disease.

Animals↗
PubMed1995

Evaluation of the SA locus in human hypertension.

The SA gene is expressed at 10-fold greater levels in the kidney of the spontaneously hypertensive rat compared with the normotensive Wistar-Kyoto rat. The gene is linked to blood pressure levels in a number of crosses involving the spontaneously hypertensive rat and other strains of genetically hypertensive rats. To assess its role in human hypertension, a human SA cDNA was cloned from a liver library. The cDNA was 1513 bp in length and exhibited a high identity with the published rat SA cDNA sequence in the coding region. A microsatellite marker was developed from a yeast artificial chromosome clone containing SA and mapped by linkage to human chromosome 16p13.11-12.3. Polymerase chain reaction amplification of human genomic DNA revealed two introns located in the SA gene, one of which contains a frequent polymorphism due to a single nucleotide substitution (cytosine to thymidine at residue 79 of the intron). Association and linkage studies in a large sample of hypertensive patients, normotensive control subjects, and multiplex sibships with these markers and other microsatellites in close proximity to SA revealed no evidence favoring involvement of the gene in the disease in humans. The methodology used in this study can be applied to the evaluation of other novel candidate genes obtained from investigations of experimental models of hereditary hypertension.

Adult↗
PubMed1995

Influence of angiotensin II type 1 receptor polymorphism on aortic stiffness in never-treated hypertensive patients.

Several clinical and experimental studies have suggested a significant role of angiotensin II in the development of alterations of small and large arteries. The present study was designed to assess the contribution of polymorphism (corresponding to an A1166-->C transversion) of the angiotensin II type 1 receptor (AT1) gene to aortic stiffness. One hundred thirty-four never-treated hypertensive patients were included in the study. Aortic distensibility was evaluated by measuring carotid-femoral pulse wave velocity. Age, systolic and diastolic pressure, and metabolic parameters were similar in the three genotypes. Pulse wave velocity was 11.4 +/- 2.5 m/s in AT1 AA homozygotes, 12.5 +/- 3.2 m/s in AC heterozygotes, and 14.7 +/- 4.0 m/s in CC homozygotes (P = .003, P < .001 after adjustment for age, blood pressure, and body mass index). Moreover, an interaction was found between AT1 genotype and the ratio of total to high-density lipoprotein cholesterol in terms of the development of aortic stiffness. Thus, a positive correlation was observed between the ratio of total to high-density lipoprotein cholesterol and pulse wave velocity in AC and CC (r = .42, P < .001) but not AA patients. These results suggest that the AT1 gene is involved in the development of aortic stiffness in hypertensive patients and could modulate the effects of lipids on large arteries.

Adult↗
PubMed1995

[Molecular biology and genetics of NO synthases].

Nitric oxide is synthesized by three isoenzymes widely distributed in the organism. The three genes encoding these enzymes show structural homology confirming that they are members of a protein family. Isoforms I (neuronal) and III (endothelial) are constitutive but their expression is transcriptionnaly regulated by various factors. NOS I promoter has not been studied yet, but alternative splicing around exons 9 and 10 has been described. SP1 binding on NOS III promoter is critical for its constitutive expression. NOS II isoform is inducible in a large variety of cells and seems also to be present constitutively in some tissues such as kidney. Functional studies of NOS II promoter reveal two important regions for lipopolysaccharides (- 85 à - 75) and interferon gamma (- 900 à - 975) induction. NOS III polymorphic markers allowed genetic studies which indicate that NOS III gene is not associated with human essential hypertension.

Animals↗
PubMed1994

A mutant renin gene in familial elevation of prorenin.

A case of familial elevation of plasma prorenin levels was discovered during an epidemiological survey of a Dutch population. Trypsin-activated prorenin was elevated in the 58-year-old father, his son, and one of his sisters. All family members were normotensive and had normal plasma renin activities. Exon sequencing of the renin gene of the proband and of his son after polymerase chain reaction amplification identified a point mutation in the last exon of the gene (exon 10). A cytosine to thymine transition creates a premature stop codon at position 387 resulting in a truncated form of renin with 20 amino acids deleted from the carboxyl terminus. All family members presenting high levels of plasma prorenin were heterozygous for the mutation. Expression vectors carrying normal or mutated renin cDNA were transiently transfected into AtT-20 cells to test in vitro the functional consequences of this mutation. Measurements of renin activity and pulse-chase experiments indicated that the truncated renin is inactive and not secreted from transfected cells. We hypothesize that the abnormal gene product of the mutated allele alters renin sorting and propose that plasma prorenin elevation may result from a compensatory mechanism.

Animals↗
PubMed1994

Identification of two active site residues in human angiotensin I-converting enzyme.

Angiotensin I-converting enzyme (ACE) contains two zinc-dependent catalytic domains (N and C domains) each bearing the motif HEXXH where the two histidines form two of the three amino acid zinc ligands. Sequence alignment of each ACE domain with other zinc metalloproteases, indicates a glutamate residues which putatively constitutes the third zinc ligand and an aspartate residue which may form an indirect zinc interaction. We investigated the functional roles of the glutamate and aspartate residues in the ACE C domain (Glu987 and Asp991) using a cDNA encoding an inactive N domain. We mutated Glu987 to aspartate (E987D) or valine (E987V) and Asp991 to glutamate (D991E) or alanine (D991A). Catalytically active mutants (E987D, D991E and D991A) exhibited similar Km values for hippuryl-His-Leu compared to non-mutated C domain. E987D displayed a 300-fold decrease in kcat and a 25-fold reduction in sensitivity to the ACE inhibitor trandolaprilat, whose binding is zinc-dependent. E987V was catalytically inactive and did not bind [3H]trandolaprilat. D991E and D991A exhibited a 3.8- and 22-fold decrease in kcat, respectively, and the Ki' values for trandolaprilat were increased 8- and 29-fold. These results provide strong evidence that Glu987 constitutes the third zinc ligand in the ACE C domain and suggest a role for Asp991 in positioning the C domain active site zinc ion.

Amino Acid Sequence↗
PubMed1994

Gene structure, polymorphism and mapping of the human endothelial nitric oxide synthase gene.

Endothelium-derived relaxing factor (EDRF)/nitric oxide (NO) is synthesized from L-Arginine by the endothelial, constitutive, NO synthase. No facilitate genetic studies, we have cloned the human endothelial NO synthase gene and determined its structure. The gene is composed of 26 exons, ranging from 68 to 579 bp and spans 22 kb. We determined the transcription start point using human lung mRNA. No TATA-box was found at the expected distance from the transcription start point and several consensus sequences for transcription factors, including a shear-stress responsive element were identified in the 5'-flanking region. A highly polymorphic (CA) repeat within intron 13 was studied, allowing the precise genetic mapping of the gene to chromosome 7, within a 4 cM interval delimited by genethon markers AFM199Zd4 and AFM074Xg5.

Alleles↗