Biomedical subjects

A C Larner

Publications and source records attributed to A C Larner.

59 records · Page 4Linked to original sources

PubMed1988

Selective transcription of an insulin-regulated gene in nuclear extracts of rat hepatoma cells.

We have previously shown that when H4 hepatoma cells are pretreated with insulin, plant lectins, phorbol esters, or insulin mediator, the steady state concentration of gene 33 mRNA is markedly increased. The increase in gene 33 mRNA concentration with insulin is due to an increase in the transcription rate of this gene. In the present report we demonstrate that nuclear extracts prepared from H4 hepatoma cells pretreated with insulin exhibit enhanced transcription of gene 33 RNA from a DNA template containing the cap site and 1500 bp upstream of the 33 gene. This is a stable effect of insulin on the nuclear RNA polymerase II system since it is observed in frozen and thawed nuclear extracts as well as fresh nuclear extracts.

Animals↗

PubMed1988

Insulin and a putative insulin metabolic mediator fraction from liver and muscle stimulate p33 messenger ribonucleic acid accumulation by apparently different mechanisms.

We have previously shown that in rat H4 hepatoma cells insulin enhances the nuclear transcription of p33 mRNA in a dose- and time-dependent manner, with no alteration in mRNA half-time (t1/2). Presumably, this effect is mediated by the cell surface receptor. In this report, we have investigated the effect of putative insulin mediator fractions which act to control metabolic events on p33 mRNA accumulation in these cells. Initial experiments originally demonstrated an insulin-like effect of an added putative metabolic fraction to enhance p33 mRNA concentrations. However, when the fetal calf serum supply was changed, the effect of insulin remained, but that of added mediator was no longer observed. After a series of experimental approaches designed to alter the permeability of the cell membrane, it was found that in the presence of increased Ca2+, the effect of mediator could again be observed. The present data demonstrate that the partially purified cAMP-dependent protein kinase/adenylate cyclase inhibitory putative mediator fractions from liver and muscle enhance p33 mRNA accumulation in intact H4 hepatoma cells by a mechanism that is differentiated from that of insulin. The action of the putative mediator is inhibited by cycloheximide, while the action of insulin itself is not. These results suggest that insulin may control nuclear transcription by multiple signaling mechanisms. Alternatively, the added putative metabolic mediator may not enter the cell in the presence of cycloheximide or is inactive as such within the cell and must first be converted to an active species by a step requiring protein synthesis.

Adenylyl Cyclase Inhibitors↗

PubMed1986

Transcriptional induction by interferon. New protein(s) determine the extent and length of the induction.

Interferon (IFN) treatment of human fibroblasts induces the transcription of two genes, IFN-IND-1 and -2, to a maximal level between 30 min and 2 h. Continued exposure to IFN results in a "desensitization" characterized by a return to pretreatment levels of transcription and the inability of further IFN treatment to reinduce the transcription of these genes. The imposition and the maintenance of desensitization apparently requires continuous protein synthesis because both can be reversed by cycloheximide. Thus a regulatory circuit of transcription for IFN-IND-1 and -2 includes a prompt IFN-induced transcriptional increase followed shortly by a negative regulatory response requiring a labile protein(s) that limits the duration of the increased transcriptional response.

Autoradiography↗

PubMed1984

Transcriptional induction of two genes in human cells by beta interferon.

The binding of interferons to distinct cell surface receptors leads to the induction of synthesis of several unique polypeptides and their corresponding mRNAs (1-6). We have isolated two cDNAs that are complementary to nuclear RNA whose synthesis is induced from undetectable levels to maximal rates of transcription within 30-60 min after the addition of beta interferon to human fibroblasts or to HeLa cells. These results prove that a single polypeptide can, by binding to a specific plasma membrane receptor, promptly activate the transcription of a defined set of genes.

DNA↗

PubMed1981

Alteration in the protein components of catecholamine-sensitive adenylate cyclase during maturation of rat reticulocytes.

The maturing rat reticulocyte was used as a model system in which to study developmental changes in the protein components of hormone-sensitive adenylate cyclase. Plasma membranes from rat erythrocytes display 10 to 20% of the adenylate cyclase activity and 30 to 50% of the beta-adrenergic receptors which are measured in membranes from rat reticulocytes, as noted by others. Reticulocyte membranes also display equal activities in response to (-)-isoproterenol in the presence of either GTP or GTP gamma S, whereas erythrocyte membrane adenylate cyclase is twice as active in the presence of isoproterenol plus GTP gamma S as in the presence of isoproterenol plus GTP. We have studied this system in greater detail by developing or applying independent assays for the catalytic protein (C) and the guanine nucleotide-binding regulatory protein (G/F) of adenylate cyclase. C was assayed in membranes by its intrinsic Mn2+-stimulated activity. It was also measured by reconstituting membranes with saturating amounts of GTP gamma S-activated G/F, yielding an operationally defined Vmax for the catalyst. By either assay, reticulocytes display about 3-fold greater C activity than do erythrocytes. G/F was assayed by its ability to confer GTP gamma S-stimulated activity upon C (which was supplied by membranes of cyc- S49 lymphoma cells). This assay indicates that reticulocyte membranes contain about 3 times as much G/F as do erythrocyte membranes. Cholera toxin and [32P]NAD were used to [32P]ADP-ribosylate the 45,000- and 52,000-dalton subunits of G/F. Total incorporation of 32P into these subunits decreased 3- to 4-fold with reticulocyte maturation. The ratio of label in the 52,000-dalton peptide to that in the 45,000-dalton peptide decreased from 0.29 in reticulocyte membranes to 0.14 in erythrocyte membranes. The apparently coordinate decrease in the amounts of C, G/F, and beta-adrenergic receptors suggest that the stoichiometry between these components is maintained during maturation, and may account for the decrease in adenylate cyclase in the membranes. However, the qualitative changes in responsiveness to hormones in the presence of GTP or GTP gamma S may be related to loss or proteolysis of the 52,000-dalton subunit of G/F.

Adenosine Diphosphate Ribose↗