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J F Catterall

Publications and source records attributed to J F Catterall.

At least 37 records · Page 2Linked to original sources
PubMed1990

Cell-specific expression of kidney androgen-regulated protein messenger RNA is under multihormonal control.

Kidney androgen-regulated protein (KAP) gene expression is under androgenic control in the epithelial cells of the proximal convoluted tubule in the mouse kidney. In Tfm/Y androgen receptor-deficient mice, KAP mRNA was detected by in situ hybridization in a subpopulation of these cells only in the S3 segment of the proximal tubules in the outer medulla. Treatment of Tfm/Y animals with testosterone caused a partial induction of KAP mRNA levels, while dihydrotestosterone had no effect. These data suggested that the androgen receptor-independent induction of KAP gene expression in these animals was mediated by an estrogenic metabolite of testosterone, since dihydrotestosterone cannot be aromatized to an estrogenic form. Estrogen treatment of Tfm/Y mice caused an increase in KAP gene expression similar to that observed with testosterone. However, ovariectomy of normal female mice did not eliminate KAP gene expression in the S3 cells and, in fact, resulted in a slight increase. Adrenalectomy in combination with castration had no effect on KAP mRNA levels in S3 cells. However, hypophysectomy alone completely eliminated this cell-specific component of KAP gene expression. These results indicate that KAP gene expression is subject to cell-specific regulation in different segments of the proximal tubule and that this regulation is mediated by hormones of both gonadal and pituitary origin.

Androgens↗
PubMed1990

Detection of spatially regulated gene expression by hybridization histochemistry.

Steady state measurements of kidney androgen-regulated protein (KAP) mRNA suggested that KAP gene expression was unusually sensitive to low hormone-receptor levels. Two of the criteria used to reach this conclusion involved relative insensitivity to treatment with a competitive antiandrogen and a partial androgen response of the gene in Tfm/Y androgen receptor (AR) deficient mice. These data may indicate the ability of the KAP gene to respond to an extremely low level of androgen-AR complex or that the effect of androgens is, at least in part, indirect. Hybridization in situ revealed that KAP mRNA expression was restricted to proximal tubule epithelial cells in the juxtamedullary region of castrated animals rather than throughout the cortex as in intact males. Examination of sections of kidneys from Tfm/Y mice before and after testosterone (T) treatment revealed that only the juxtamedullary tubules expressed KAP mRNA and that T increased the level of KAP mRNA in these cells. Treatment of Tfm/Y animals with other steroids showed that beta-estradiol treatment mimicked the effect of T while dihydrotestosterone (DHT) had no effect. The facts that DHT and T both stimulate cortical expression of KAP mRNA in normal animals but DHT has no effect on the juxtamedullary cells in the Tfm/Y variant may indicate that the action of T is due to an estrogenic metabolite. Castrated, hypophysectomized males exhibited no KAP gene expression, while in the presence of T, expression was observed throughout the cortex as in intact males. These results clearly indicate the involvement of pituitary hormones in KAP gene expression in the juxtamedullary tubules. These studies have shown that the regulation of KAP gene expression in the mouse kidney is much more complex than originally believed. Future studies will further investigate the roles of estrogen and specific pituitary hormones in KAP gene expression.

Androgen-Insensitivity Syndrome↗
PubMed1989

Nucleotide sequence of kidney androgen-regulated protein mRNA and its cell-specific expression in Tfm/Y mice.

Kidney androgen-regulated protein (KAP) mRNA is an abundant renal mRNA that was originally identified by comparisons of the products of in vitro translation of poly(A) RNA from animals before and after androgen stimulation. KAP mRNA is 607 nucleotides long, excluding its poly(A) segment, and encodes a protein of 13,265 mol wt. A hydrophobic N-terminal domain forms a putative signal peptide of 18 amino acids, the cleavage of which results in a 103-amino acid mature protein with a molecular size of 11,297. The protein is highly negatively charged and contains regions of clustered Pro, Glu/Asp, Ser, and Thr residues that are associated with proteins with short half-lives. KAP mRNA is unusual in that it is expressed in two distinct regions of the kidney under different hormonal treatments. It is expressed throughout the cortex in the epithelial lining of the proximal tubules in response to androgen stimulation. After castration, only tubules in the outer stripe of the medulla express KAP mRNA. The androgen receptor-deficient Tfm/Y mutant strain exhibits KAP mRNA induction only in this juxtamedullary region after testosterone treatment. Expression of KAP mRNA in these cells is responsible for the relatively high basal levels of KAP mRNA in female and castrated male animals, and induction in these cells occurs by an androgen receptor (AR)-independent mechanism.

Animals↗
PubMed1988

Differential regulation of gene expression during macrophage activation with a polyribonucleotide. The role of endogenously derived IFN.

The activation of macrophages by exposure to the polyribonucleotide, poly [I:C], is accompanied by a large stimulation of the synthesis of the C components factor B and C3, and a concomitant inhibition of the synthesis of the lysosomal enzyme beta-glucuronidase. Northern blot analysis of poly [A+] RNA extracted from poly [I:C]-stimulated cells revealed that the changes in the synthesis of factor B and C3 were related to changes in the levels of their respective mRNA and hence the expression of these proteins appeared to be regulated at a pre-translational level. The down-regulation of the synthesis of beta-glucuronidase appeared to be regulated at both translational and pre-translational levels. In view of the proposed role of macrophage-derived IFN in the regulation of macrophage activation, we investigated the possible role of IFN-alpha/beta in the regulation of the synthesis of factor B, C3, and beta-glucuronidase. Exposure of macrophages to mouse IFN-alpha and IFN-beta induced limited changes in the synthesis of factor B, C3, and beta-glucuronidase. However, pretreatment of macrophages with only 500 U/ml of IFN-beta primed the cells thereby increasing their sensitivity to poly [I:C]. IFN-alpha was less effective as a priming agent. When macrophages were exposed to poly [I:C] in the presence of an anti-mouse IFN-alpha/beta antiserum, the changes in the synthesis of factor B, C3, and beta-glucuronidase were partially inhibited. Collectively, these data indicate first, that exposure of mouse bone marrow-derived macrophages to poly [I:C] differentially regulates the expression of the products of the genes for factor B, C3, and beta-glucuronidase. Second, IFN-alpha and IFN-beta prime macrophages to increase the sensitivity of macrophages to poly [I:C]. Third, in the absence of exogenous IFN, macrophage-derived IFN appears to participate in priming the cells in an autocrine or paracrine fashion.

Animals↗
PubMed1988

Genomic organization and sequence of the Gus-s alpha allele of the murine beta-glucuronidase gene.

The Gus-s alpha allele of the mouse beta-glucuronidase gene exhibits a high degree of inducibility by androgens due to its linkage with the Gus-r alpha regulatory locus. We isolated Gus-s alpha on a 28-kilobase pair fragment of mouse chromosome 5 and found that it contains 12 exons and 11 intervening sequences spanning 14 kilobase pairs of this genomic segment. The mRNA cap site was identified by ribonuclease protection and primer extension analyses which revealed an unusually short 5' noncoding sequence of 12 nucleotides. Proximal regulatory sequences in the 5'-flanking DNA and the complete sequence of the Gus-s alpha mRNA transcript were also determined. Comparison of the amino acid sequence determined from the Gus-s alpha nucleotide sequence with that of human beta-glucuronidase indicated that the two human mRNA species differ due to alternate splicing of an exon homologous to exon 6 of the mouse gene.

Alleles↗
PubMed1987

Unique patterns of androgen regulation of the expression of two genes in murine kidney.

The kidneys of androgen stimulated mice exhibit a hypertrophic response but no hyperplasia or concomitant DNA replication. Androgens increase the expression of several genes in mouse kidney. The response of the beta-glucuronidase gene to testosterone in this tissue is characterized by a 1-2 day lag and relatively slow induction kinetics. The gene coding for kidney androgen-regulated protein (KAP) exhibits quite a different response to the hormone when compared on the basis of initial response to a given dose, dose required to produce maximal response, and apparent sensitivity to low levels of androgen-receptor complexes in renal nuclei. The analysis of the accumulation of the mRNAs produced by these two genes suggests that gene-specific differential sensitivity to androgen receptor complexes governs the development of the cellular male phenotype in this tissue.

Androgens↗
PubMed1987

Mouse kidney androgen-regulated protein messenger ribonucleic acid is expressed in the proximal convoluted tubules.

Kidney androgen-regulated protein (KAP) mRNA represents the most abundant [approximately 4% of the total poly(A) RNA] mRNA species that is induced by androgens in the mouse kidney. Comparison of the expression of several androgen-regulated mRNAs in this tissue, revealed that the mRNAs were differentially regulated by the hormone. KAP mRNA exhibited unusual sensitivity to low concentrations of the androgen-receptor complex. Because of its unusual characteristics, it was of interest to determine in what cells of the kidney KAP mRNA was being produced. Using the technique of in situ hybridization with single stranded RNA probes, we have identified the epithelial cells of the renal proximal convoluted tubules as the site of synthesis of KAP mRNA. Interestingly, only a subpopulation of these cells, those located in the juxtamedullary region of the renal cortex, contain KAP mRNA in castrated males. After androgen treatment, cortical proximal tubules are also induced to express KAP mRNA. These results suggest that two types of response to androgens occur in these cells. One is the progressive increase of KAP gene expression in the juxtamedullary region while the other involves recruitment of new cells in the cortical proximal tubules to synthesize KAP mRNA.

Androgens↗
PubMed1986

Genetic regulation of androgen-induced accumulation of mouse renal beta-glucuronidase messenger ribonucleic acid.

Mouse kidney beta-glucuronidase is induced by androgens in a receptor-dependent fashion. Genetic regulatory mutations have been described which govern this response. In mice carrying the Gus-ra allele (A/J), the induction of enzyme activity is 3-5 times greater than in animals with the Gus-rb allele (C57BL/6J). To study this hormonal and genetic control at the molecular level, we measured changes in beta-glucuronidase mRNA concentrations in these two mouse strains using cloned cDNA. The specificity of the regulation was assessed by following changes in the concentration of kidney androgen-regulated protein (KAP) mRNA, which is also under androgen control in mouse kidney. Female mice were treated with Silastic implants that released 120 micrograms testosterone/day over a 20-day time course. Induction of specific mRNA was analyzed by either Northern blot hybridization or a more quantitative assay in which renal poly(A) RNA was covalently bound to aminophenylthioether paper discs. The induction of beta-glucuronidase mRNA was about 10-fold higher in the strain carrying the Gus-ra regulatory allele, indicating that the Gus-r locus controls the beta-glucuronidase structural gene (Gus-s) at the level of mRNA accumulation. Regulation by Gus-r was specific for beta-glucuronidase mRNA as kidney androgen-regulated protein mRNA accumulation did not differ between these two strains of mice after androgen treatment.

Animals↗
PubMed1983

Detection of early changes in androgen-induced mouse renal beta-glucuronidase messenger ribonucleic acid using cloned complementary deoxyribonucleic acid.

beta-Glucuronidase mRNA was purified from androgen-induced mouse kidney by immunoadsorption of polysomes to protein A-Sepharose. Cell-free translation of mRNA isolated from the protein A bound RNA followed by immunoprecipitation revealed that beta-glucuronidase mRNA represented approximately 2% of the purified mRNA fraction. This mRNA preparation was used to produce complementary DNA clones by recombination with pBR322. Clones containing sequences that were enriched during the purification procedure were selected by differential colony hybridization. These were further screened for homology with beta-glucuronidase mRNA by hybrid-selected translation. A beta-glucuronidase cDNA clone, designated pGUS7, was identified by these criteria. With this plasmid, the abundance of beta-glucuronidase mRNA in total poly(A) mRNA from androgen-induced mouse kidney was estimated to be less than 0.04%. The beta-glucuronidase cDNA plasmid hybridized to a mRNA of 2.6 kb in length, which was induced in an androgen receptor dependent fashion over a time course of 21 days. Treatment of female mice with a single dose of testosterone (10 mg) revealed that beta-glucuronidase mRNA concentration begins to increase between 12 and 24 h after hormone administration.

Androgens↗
PubMed1983

Cloning of cDNA encoding the pre-beta subunit of mouse thyrotropin.

Double-stranded cDNA was synthesized from sucrose gradient-purified poly(A)+ mRNA from a mouse thyrotropic tumor, inserted into the Pst I site of plasmid pBR322 by using poly(dC) . poly(dG) homopolymeric extensions, and cloned in Escherichia coli RRI. Plasmids containing cDNA sequences coding for the beta subunit of thyrotropin (TSH) were identified by cell-free translation of hybrid-selected mRNA and immunoprecipitation with specific antibody to TSH beta subunit. Determination of the nucleotide sequence of one cDNA, 595 base pairs in length, allowed us to deduce the entire amino acid sequence of the mouse TSH beta subunit. The pre-beta subunit contains a 20-amino acid amino-terminal signal sequence followed by a 118-amino acid mature TSH beta subunit. There is 85-90% homology in amino acid sequence between mouse TSH beta subunit and subunits from man, pig, and cow; however, the mouse subunit contains an additional 5 or 6 amino acids at its carboxyl terminus compared to the bovine or human and pig subunits, respectively. TSH beta-subunit mRNA from mouse thyrotropic tumor was estimated to be 750 nucleotides in length by hybridization with labeled TSH beta-subunit cDNA.

Amino Acid Sequence↗
PubMed1981

Complete nucleotide sequence of the chicken chromosomal ovalbumin gene and its biological significance.

The nucleotide sequence of the entire chicken chromosomal ovalbumin gene has been determined. The gene is 7564 nucleotides in length to code for a mature messenger RNA of 1872 nucleotides. Comparison of the sequence at the 5'-terminal region of the gene with that reported by others has revealed multiple polymorphic nucleotides in the structural, intervening, and flanking DNA sequences. Some of the polymorphic sites occur at positions very close to splice junctions or the eucaryotic promoter sequence, yet apparently have little or no effect on the expression of this gene. The heptanucleotide promoter sequence TATATAT present in the 5'-flanking region of the ovalbumin gene does not occur within the confines of the gene. Nevertheless, multiple Hogness box sequences similar to those found in other eucaryotic genes were delineated within the boundaries of the gene. These internal Hogness box sequences are not used for transcription initiation. Similarly, the hexanucleotide sequence AATAAA common to all eucaryotic messenger RNAs at the 3'-untranslated region occurs seven additional times within the ovalbumin gene. These sites are not used for transcription termination or polyadenylation. Thus, although these sequences may play important roles in the initiation or termination of gene transcripts as well as polyadenylation of the transcripts, the specificity for such biological functions must not reside within these sequences alone. Furthermore, sequences complementary to the highly conserved rat U1 small nuclear RNA have been found throughout the gene. Many of these regions of complementarity occur in the structural sequences. If the small nuclear RNA does play a role in splicing, the specificity must be provided also by other as yet undefined components.

Animals↗