Hormonal inducibility of rat α2u globulin genes in transfected mouse cells

Kurtz, David T.

doi:10.1038/291629a0

Cited by 137 publications

(45 citation statements)

References 17 publications

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“…Two rat a2u-globulin genes introduced by co-transfection with the herpes virus-thymidine kinase gene in the tk-L cells were transcribed and expressed as secreted a2u-globulin. Both the mRNA and the protein levels rose in response to the steroid (Kurtz, 1981). Similar results in the same system were obtained with the rat (Moore et al, 1982) and human somatotropin genes and showed that DNA sequences required for induction reside within 500bp of 5'-flanking DNA (Robins et al, 1982).…”

Section: Glucocorticoids Modulate Mrna Concentrationsupporting

confidence: 69%

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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Section: Glucocorticoids Modulate Mrna Concentrationsupporting

confidence: 69%

Control of gene expression by glucocorticoid hormones

Rousseau¹

1984

Biochemical Journal

142

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“…Using these techniques, investigators have introduced several eucaryotic genes into foreign cells, and their expression has been demonstrated at the nucleic acid level, protein level, or both (Mulligan et al, 1979;Wigler et al, 1979;DeRobertis and Olson, 1979;Capecchi, 1980;Grosschedl and Birnstiel, 1980;Lai et al, 1980). Furthermore, there are recent reports of regulation of gene expression in these systems (Kurtz, 1981;Buetti and Diggelmann, 1981;Hynes et al, 1981). For other genes, we anticipate that expression or regulation may be difficult to demonstrate because the appropriate cell type may not be amenable to analysis in culture, or developmental programming may be essential.…”

Section: Introductionmentioning

confidence: 99%

Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs

Brinster

Chen

Trumbauer

et al. 1981

Cell

636

223

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SummaryA plasmid, designated pMK, containing the structural gene for thymidine kinase from herpes simplex virus (HSV) fused to the promoter/regulatory region of the mouse metallothionein-I gene, was injected into the pronucleus of fertilized one-cell mouse eggs; the eggs were subsequently reimplanted into the oviducts of pseudopregnant mice. The first experiment produced 19 offspring, one of which expressed high levels of HSV thymidine kinase activity in the liver and kidney. pMK DNA sequences were detected in equal amounts in several tissues of the expressing mouse as well as in three mice that did not express HSV thymidine kinase activity. In all cases, several copies of the pMK plasmid were tandemly duplicated and integrated into mouse DNA. It appears as though multiple copies of the intact plasmid were fused by homologous recombination either before or after integration at a single site in the mouse genome. The overall efficiency of obtaining somatic expression of thymidine kinase in experiments performed to date is about 10% (4/41), and twice this number have integrated pMK DNA. This procedure not only provides a means of introducing new genes into mice, but it will also be a valuable system for studying tissue-specific regulation of gene expression.

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“…In FaC1 a portion of the RNA was lost, which accounts for the decreased signal. (29)(30)(31)(32), and a few of these appear to be transcriptionally activated given the proper induction signal (33,34), cell background plays an important role in the regulation of tissue-specific gene expression (33,(35)(36)(37) As shown in other systems (38,39), the orientation of the insert is probably not a major factor in the regulation of expression for the chicken actin genes, because the genes are being transcribed from their respective promotors, as judged by primer extensions, and are flanked on their 5' and 3' sides by a similar length of sequence. The level of expression may, however, depend on orientation, and this is under study.…”

Section: Introduction Of the Chicken Actin Genes Into Myogenic C2mentioning

confidence: 99%

Expression and regulation of chicken actin genes introduced into mouse myogenic and nonmyogenic cells.

Seiler-Tuyns¹,

Eldridge²,

Paterson³

1984

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

105

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We have introduced the chicken genes for cytoplasmic 13-actin', cardiac a-actin, and skeletal a-actin into :2 cells, a murine myogenic cell line, and into L cells by using the simian virus 40-derived vector PSV2-gpt. In each selection, the entire population of transformed cells was analyzed for the expression and regulation of the actin genes by nuclease S1 assay and primer extension. This was compared to the expression of the vector marker Eco-gpt. The .8-actin gene is transcribed accurately and efficiently both in L-cells and in undifferentiated C2 cells.'In fused C2 cells, (8-actin transcripts decrease significantly in parallel with the endogenous level of mouse f-actin mRNA. Eco-gpt RNA levels remain essentially constant during myogenesis. The a-actin genes are correctly expressed at low levels in L cells but at significantly higher levels in the C2 cell background. Unlike the endogenous mouse a-actin gene, this level of expression does not change measurably with inyogenesis. The skeletal a-actin gene is expressed poorly in pre-and post-fusion C2 cells, displaying no induction with differentiation. These results suggest that the tissue specificity of expression is maintained but the'pattern of gene regulation for the sarcomeric actins is not. Factors in addition to the sequences flanking these genes are important for modulating gene expression during development. The decrease in the levels of g3-actin RNA during C2 cell differentiation provides a model system in which to'study gene repression during development.

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Hormonal inducibility of rat α2u globulin genes in transfected mouse cells

Cited by 137 publications

References 17 publications

Control of gene expression by glucocorticoid hormones

Control of gene expression by glucocorticoid hormones

Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs

Expression and regulation of chicken actin genes introduced into mouse myogenic and nonmyogenic cells.

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