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F Poulat

Publications and source records attributed to F Poulat.

28 records · Page 2Linked to original sources
PubMed1994

The human testis determining factor SRY: a new member of the HMG box protein family.

The product of the sex-determining gene SRY is a member of the HMG box containing protein superfamily. The HMG box is a DNA-binding domain of about 80 amino acids shared by many proteins with diverse functions. It seems that the functions of the full length protein are restricted to the HMG box but their molecular basis remains to be determined. We have summarized here the properties of this binding domain described so far in the literature and, using a synthetic peptide mimicking the DNA binding domain (SRY80), we have confirmed the existence of DNA minor groove contacts with this domain. Using intrinsic fluorescence of the tryptophane, the interaction between SRY80 and the putative target sequence AACAAAT was also quantified. In conclusion, we also consider the possible putative action of SRY to fulfill its role in sex determination.

Amino Acid Sequence↗
PubMed1994

[Human testicular determining factor: from the gene to protein].

Since its cloning in 1990, the human SRY gene has been formally identified with the testis determining factor. The SRY gene encodes a 204 amino acid protein of the High Mobility Group family. Its ability to bind DNA, to bend DNA or to be translocated into the nuclear compartment of the cell have now been established. However, neither its mode of action nor the description of target genes have been described so far, and are the topics of many studies.

Amino Acid Sequence↗
PubMed1993

Distinct molecular origins for Denys-Drash and Frasier syndromes.

The direct involvement of the Wilm's tumor suppressor gene (WT1) in Denys-Drash syndrome through mutations within exons 8 or 9 has recently been established. The absence of such alterations in three patients with Frasier syndrome provides a molecular basis for distinguishing these two syndromes that are associated with streak gonads, pseudohermaphroditism and renal failure.

Base Sequence↗
PubMed1992

Synthesis of a large peptide mimicking the DNA binding properties of the sex determining protein, SRY.

The sex determining protein, SRY, has been recently described as containing a DNA binding motif, also called the SRY box. This 80 amino acid box was synthesized using the continuous flow solid-phase technique. The product was then purified and tested according to such diverse criteria as its intrinsic structure or its biological activity (DNA binding capacity), and compared to the full-length protein. The data indicate that the peptide is relevant for the properties described so far for the protein.

Amino Acid Sequence↗
PubMed1992

Molecular analysis of the sex-determining region from the Y chromosome in two patients with Frasier syndrome.

In the Frasier syndrome there is an association between XY gonadal dysgenesis and chronic renal failure. Owing to an observed sex reversal, the Y chromosomes of two girls with this syndrome have been analyzed. Using molecular-biology techniques, no major alterations of the known sex-determining area of the Y chromosome were found. Furthermore, the sequence did not reveal impairment of the recently described testis-determining factor SRY. These data suggest that in the Frasier syndrome, XY sex reversal and renal failure could be the result of either faulty gene(s) located downstream in the sex differentiation pathway during embryogenesis, or impaired SRY regulation. Preliminary results on the Wilms' tumor suppressor gene WT1, a candidate for acting downstream to SRY, are also provided.

Adolescent↗
PubMed1991

Pharmacological characterization of the quisqualate receptor coupled to phospholipase C (Qp) in striatal neurons.

A detailed pharmacological characterization of the quisqualate (QA) receptor coupled to phospholipase C (Qp) was performed in striatal neurons. The experiments were carried out in the presence of the ionotropic antagonists MK-801 (1 microM) and 6-cyano-7-nitroquinoxaline-2,3-dione (30 microM), concentrations that block N-methyl-D-aspartate (NMDA) or alpha-amino-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in these cells. QA, ibotenate and trans-1-aminocyclopentyl-1,3-dicarboxylate (ACPD) evoked dose-dependent inositol phosphate formations with EC50 values of 0.3, 6.7 and 29 microM, respectively. QA and ibotenate had the same maximal effect (295.7 +/- 17.9% of basal, n = 6) whereas the efficacy of ACPD was somewhat lower (70.2 +/- 8.9% of the maximal quisqualate effect, n = 4). The QA-, ibotenate- and ACPD-induced maximal effects were not additive, and the inositol phosphate formations induced by high concentrations of L-aspartate (L-ASP), AMPA, kainate (KA) and domoate (DO) (100 microM or higher) were also not additive. The inositol phosphate responses induced by all these agonists were totally blocked by the phorbol ester phorbol 12,13-dibutyrate (PdBu), but not by atropine or prazosin suggesting that all these substances were able to stimulate the Qp excitatory amino acid receptor in striatal neurons. Of the excitatory amino acid receptor antagonists tested, only D,L-2-amino-3-phosphonopropionate (D,L-AP3) inhibited QA-induced InsP formation in a competitive manner (mean pKi = 4.45 +/- 0.43, n = 4). However, this drug was also a partial agonist of the Qp receptor since it stimulated the inositol phosphate formation. We found that D,L-AP3 also inhibited NMDA-induced calcium increase, in a competitive manner (mean pIC50 = 4.34 +/- 0.22, n = 8, and mean pKi = 3.7 +/- 0.11, n = 5). The Qp excitatory amino acid receptor in striatal neurons therefore closely resembles Qp receptors with high potency for agonists as described in striatal and retinal slices and synaptoneurosomes, and has several pharmacological differences compared to the Qp receptors which have low potency for agonists described in hippocampal and cortical slices, cerebellar granule cells, astrocytes and rat brain mRNA-injected oocytes.

2-Aminoadipic Acid↗
PubMed1991

[Determination of sex and Y chromosome].

Years of speculations about the nature of the elusive testis determining factor (TDF) of the Y chromosome have ended last year. A gene named SRY satisfies many criteria expected of the testis determining gene, and gives us a basis to understand molecular mechanisms of the testis differentiation. The different steps which gave rise to SRY cloning are described.

DNA-Binding Proteins↗
PubMed1990

(trans)-1-amino-cyclopentyl-1,3-dicarboxylate stimulates quisqualate phosphoinositide-coupled receptors but not ionotropic glutamate receptors in striatal neurons and Xenopus oocytes.

The effects of a novel glutamate analogue, (trans)-1-amino-cyclopentyl-1,3-dicarboxylate (ACPD), have been tested in striatal neurons in primary culture and in Xenopus oocytes injected with rat brain RNA. Both systems have been previously shown to contain well characterized metabotropic receptors coupled to phospholipase C (Qp), as well as ionotropic glutamate receptors. In striatal neurons, ACPD stimulated inositol phosphate (InsP) accumulation (EC50 = 9.7 +/- 2.5 microM; maximal effect, 184.7 +/- 11.6% of basal accumulation). This effect of ACPD was likely to be mediated by Qp receptors, because maximal ACPD and quisqualate-induced InsP formation were not additive. In contrast, the effects of ACPD and norepinephrine on InsP formation were additive. ACPD-induced InsP formation was not antagonised by antagonists of muscarinic and alpha 1-adrenergic receptors (1 microM atropine and 0.1 microM prazosin, respectively). In Xenopus oocytes, ACPD and quisqualate induced an oscillatory increase of a Ca2(+)-dependent chloride conductance, which is characteristic of the activation of phospholipase C-coupled receptors in this model. The specificity of ACPD on Qp receptors was demonstrated by testing the effect of this drug on quisqualate/kainate as well as on N-methyl-D-aspartate ionotropic receptors. In striatal neurons, the activation of quisqualate/kainate and N-methyl-D-aspartate receptors was tested by measurement of [3H]-gamma-aminobutyric acid release and by electrophysiological recordings using the patch-clamp technique. At concentrations as high as 1 mM, ACPD was inactive on these inotropic receptors, either as agonist or as antagonist. In conclusion, ACPD appeared to be a highly specific agonist of Qp receptors, with no activity on ionotropic glutamate receptors. It will be a useful drug to study the physiological properties of Qp receptors in vertebrate brains.

Amino Acids↗