Identification of glucocorticoid-induced genes in rat hepatoma cells by isolation of cloned cDNA sequences.

Feinberg, Ronald F.; Sun, Lee K.; Ordahl, Charles P.; Frankel, Fred R.

doi:10.1073/pnas.80.16.5042

Cited by 16 publications

(6 citation statements)

References 36 publications

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“…Glucocorticoid treatment of HTC cells leads to a dramatic increase in the production of AGP and its mRNA (5)(6)(7). Since the increased accumulation of AGP mRNA is blocked by cycloheximide (5-7), we performed experiments to examine the mechanism by which this indirect (secondary) effect of glucocorticoids occurs.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Glucocorticoid-mediated induction of alpha 1-acid glycoprotein: evidence for hormone-regulated RNA processing.

Vannice¹,

Taylor²,

Ringold³

1984

Proc. Natl. Acad. Sci. U.S.A.

196

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We have studied the glucocorticoid-mediated accumulation of a,-acid glycoprotein (AGP) knRNA in HTC rat hepatoma cells. In contrast to the well-characterized primary response of mouse mammary tumor vills, in viro transcription assays in isolated nuclei show that the rate of transcription of the AGP gene is high even in the absence of hormone. Despite the constitutive transcription of the AGP gene, no detectable AGP RNA can be found in either the cytoplasm or the nuclei of untreated cells. Previous experiments have shown that the glucocorticoid induction of AGP RNA requires ongoing protein synthesis. In conjunction with the present study, our data suggest that glucocot-ticoids stimulate accumulation of AGP RNA by inducing an RNA processing factor that allows production of stable transcripts.The acute phase reactant a,-acid glycoprotein (AGP)K also called orosomucoid, is one of several plasma proteins synthesized by the liver in response to various stressful stimuli. Physical trauma such as surgery or wounding, bacterial infection, or nonspecific inflammatory stimuli such as subcutaneous injection of turpentine elicit the so-called acutephase response (for review, see ref. 1). In addition to AGP, whose physiological function remains obscure, a variety of protease inhibitors (e.g., a1-anti-trypsin) and iron scavengers (e.g., haptoglobin and hemopexin) are also induced. However, the mechanisms by which acute-phase reactants are induced are poorly understood. Macrophage factors, in particular interleukin I (2), and glucocorticoids (3, 4) have both been implicated in induction of some acute-phase reactant proteins. We (5), Baumann et al. (6), and Feinberg et al. (7) have recently shown that AGP and its mRNA are induced several hundred-fold by glucocorticoids in the rat hepatoma cell line HTC.Glucocorticoids, and steroid hormones in general, act by altering the rates of transcription of specific genes. The model for steroid hormone action, originally proposed by Jensen et al. (8) The gene encoding AGP, however, appears to be regulated in quite a different manner. The several hundred-fold increase in AGP RNA observed in glucocorticoid-treated HTC cells is dependent on ongoing protein synthesis (5-7); this is in contrast to the induction of MMTV RNA, which proceeds normally in the absence of protein synthesis (16,17). Thus, the induction of AGP RNA by glucocorticoids is a secondary action of the hormone, presumably mediated by a protein that is induced by direct action of the glucocorticoid-receptor complex. In this manuscript, we describe experiments showing that the induction of AGP RNA occurs at a post-transcriptional level. Our data implicate a glucocorticoid-regulated RNA processing system in the production of AGP RNA. MATERIALS AND METHODSCells and Cell Culture. The cell lines used in these studies were derived from the rat hepatoma cell line HTC (18). JZ.1 and MSC1 are HTC-derived cell lines containing 1 and -10 MMTV proviral copies, respectively. They are described in more detail elsewhere (19). These ce...

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Section: Discussionmentioning

confidence: 99%

“…Macrophage factors, in particular interleukin I (2), and glucocorticoids (3,4) have both been implicated in induction of some acute-phase reactant proteins. We (5), Baumann et al (6), and Feinberg et al (7) have recently shown that AGP and its mRNA are induced several hundred-fold by glucocorticoids in the rat hepatoma cell line HTC.…”

mentioning

confidence: 99%

Glucocorticoid-mediated induction of alpha 1-acid glycoprotein: evidence for hormone-regulated RNA processing.

Vannice¹,

Taylor²,

Ringold³

1984

Proc. Natl. Acad. Sci. U.S.A.

196

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“…Cycloheximide abolishes in rat liver cells the induction of a2u-globulin (Chen & Feigelson, 1979) and of phosphoenolpyruvate carboxykinase (Iynedjian & Jacot, 1981) mRNA; it reduces the accumulation of a 1-acid glycoprotein mRNA (Feinberg et al, 1983). In rat glial cells, the glucocorticoid-induced accumulation of mRNA for glycerol phosphate dehydrogenase-is inhibited by cycloheximide (McGinnis & de Vellis, 1978 (Firestone et al, 1982).…”

Section: Glucocorticoids Modulate Mrna Concentrationmentioning

confidence: 99%

“…In constructs containing only one LTR, glucocorticoids both stimulated thymidine kinase transcription and allowed initiation at the LTR cap site, as if the hormone could substitute for the missing, second, LTR (Groner et al, 1983b). The complexity of glucocorticoid action is further illustrated by the facts that it superimposes to that of viral (SV40 or murine sarcoma) enhancers (Majors & Varmus, 1983), and that it appears to stimulate transcription of repetitive sequences homologous to the Alu family (Feinberg et al, 1983) (Alu sequences are short, repetitive, stretches of DNA that are complementary to sequences found in small cytoplasmic RNAs such as the 7 S RNA component of the 'signal recognition particles').…”

Section: Transcriptional and Post-transcriptional Effectsmentioning

confidence: 99%

Control of gene expression by glucocorticoid hormones

Rousseau¹

1984

Biochemical Journal

142

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Glucocorticoids control the expression of a small number of transcriptionally active genes by increasing or decreasing mRNA concentration. Either effect can result from a transcriptional or a post-transcriptional mechanism. Induction of mouse mammary tumour virus RNA results from a stimulation of transcription initiation and depends on the presence of defined regions in proviral DNA. These regions bind the glucocorticoid receptor and behave functionally as proto-enhancers. Glucocorticoid-inducible genes can retain their sensitivity to the hormone after transfer to a heterologous cell by transfection techniques. Non-inducible genes can become inducible when linked to the promoter region of an inducible gene. The mechanisms by which the receptor-steroid complex stimulates or inhibits transcription or influences mRNA stability are unknown. Receptor binding to nucleic acids appears to be a necessary but not sufficient condition. It is likely that the receptor also interacts with chromatin proteins. This might lead to a catalytic modification of these proteins, resulting in a modulation of gene expression. Development of glucocorticoid-sensitive, biochemically defined, cell-free transcription systems should provide a tool to delineate the molecular determinants of this essential regulatory mechanism.

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“…Double-stranded cDNA was synthesized from 5 ,ug of poly(A)+ mRNA isolated from ts13 cells that had been serum-stimulated for 6 hr at the permissive temperature (16). After digestion with Sau3A1, the cDNA was ligated into the BamHI site of pBR322 (17). Recombinant plasmids transformed into E. coli strain HB101, which were ampicillin resistant and tetracycline sensitive, were replica-plated from 96-well microtiter plates onto nitrocellulose paper in duplicate.…”

Section: Methodsmentioning

confidence: 99%

Cell-cycle-specific cDNAs from mammalian cells temperature sensitive for growth.

Hirschhorn¹,

Aller²,

Yuan³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

181

135

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A library of double-stranded cDNA was constructed from ts13 cells, a Gl-specific temperature-sensitive hamster cell line. The cDNAs, cloned into pBR322, were prepared from poly(A)+ mRNA isolated from ts13 cells 6 hr after serum stimulation at the permissive temperature of 34°C.Differential screening of the library with G -specific and Gospecific single-stranded cDNA probes prepared from the same cells identified five cDNA clones whose sequences were preferentially expressed in G1. (2), is a mutant of RNA polymerase II (3-6). In addition, the progression of cells in culture through G1 can be inhibited by the intracellular microinjection of a-amanitin (5), a drug that at low concentrations is known to have one and only one specific site of action, the large subunit of RNA polymerase II (7). These studies conclusively showed that a functional RNA polymerase II is an absolute requirement for the transition of cells from a resting to a growing state. This requirement justifies the search for genes transcribed by RNA polymerase II that control the transition from Go to G1 to S. As a first step in this direction, we have begun a search for genes whose expression is specifically increased in G1. A cDNA library was constructed from poly(A)+ mRNA isolated from serumstimulated ts13 cells that are a G1-specific temperature-sensitive mutant of the cell cycle (2,8). Genes that were preferentially expressed in G1 were identified by differential hybridization. The combination of molecular biology techniques with the use of Gl-specific temperature-sensitive mutants should ultimately allow us to select among the preferentially expressed genes those that play a major role in cell-cycle progression. This initial report describes the identification and characterization of five cDNA clones that are preferentially expressed in G1 at the permissive temperature yet are differently affected by the temperature-sensitive block at the restrictive temperature. MATERIALS AND METHODSCell Lines and Culture Conditions. The cell lines ts13 and tsAF8 are G1-specific temperature-sensitive mutants originally isolated from baby hamster kidney (BHK) cells (2, 8). Both lines were maintained under culture conditions described in detail (9, 10). The permissive temperature for both cell lines was 34WC, and the nonpermissive temperature was 39.60C for ts13 and 40'C for tsAF8. Cells were made quiescent by serum deprivation [i.e., maintained 48-50 hr in Dulbecco's minimal essential medium (DME medium) supplemented with 0.5% calf serum]. Quiescent populations were stimulated with fresh DME medium containing 15% fetal calf serum. The entry of cells into DNA synthesis was routinely monitored by continuous labeling with [3H]thymidine (6.7 Ci/mmol; 0.5 ,Ci/ml; 1 Ci = 37 GBq; New England Nuclear) followed by autoradiography.Enzymes. All restriction endonucleases were purchased from Bethesda Research Laboratories or New England Biolabs and were used according to the manufacturers' directions. Avian myeloblastosis virus reverse transcriptase was provided by...

show abstract

Identification of glucocorticoid-induced genes in rat hepatoma cells by isolation of cloned cDNA sequences.

Cited by 16 publications

References 36 publications

Glucocorticoid-mediated induction of alpha 1-acid glycoprotein: evidence for hormone-regulated RNA processing.

Glucocorticoid-mediated induction of alpha 1-acid glycoprotein: evidence for hormone-regulated RNA processing.

Control of gene expression by glucocorticoid hormones

Cell-cycle-specific cDNAs from mammalian cells temperature sensitive for growth.

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