Hormonal regulation of chimeric genes containing the phosphoenolpyruvate carboxykinase promoter regulatory region in hepatoma cells infected by murine retroviruses.

M. Hatzoglou, Edwards Park, A. Wynshaw-Boris, H. L. Kaung, R. W. Hanson

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Hepatoma cells were infected with replication-incompetent murine retroviruses containing the selectable gene for amino-3'-glycosyl phosphotransferase (neo) and/or the nonselectable gene for bovine growth hormone (bGH). Expression of these genes was controlled by the promoter regulatory region of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK) from the rat, which contains hormone and tissue-specific regulatory elements. Expression of the transduced PEPCK-neo gene was stimulated by Bt2cAMP and glucocorticoids and inhibited by insulin. The amount of RNA which initiated within the retroviral 5' long terminal repeat (5' LTR) was inhibited when internal promoters were present in the retroviral vector. When no internal promoter was present, expression from the 5' LTR was higher and stimulated by glucocorticoids, due to the presence of a glucocorticoid regulatory element in the 5' LTR. Infection of cells with retroviruses altered the basal expression and hormonal regulation of the endogenous PEPCK gene, but had no effect on the expression of the tyrosine aminotransferase gene, which is regulated in a similar manner by cAMP and glucocorticoids. A segment of the PEPCK promoter acted as a hormonally regulated enhancer, bringing the SV40 early promoter under the control of Bt2cAMP. A second, nonselectable gene (PEPCK-bGH), contained in the retroviral vector together with PEPCK-neo, was expressed and regulated appropriately when introduced into hepatoma cells. The proviruses were initially integrated randomly into the host cell genome, but after prolonged selection for expression of the transduced PEPCK-neo gene, cells were selected which contain a predominant site(s) of integration. Among populations of cells, however, the predominant site(s) of proviral integration was different. The selection of cells with a specific site of integration from a population was accelerated by the presence of PEPCK promoter sequences in the provirus. Despite the need to better characterize their effects on the host cell, retroviruses appear to be versatile tools for the specific introduction of regulated genes into cells.

Original languageEnglish (US)
Pages (from-to)17798-17808
Number of pages11
JournalThe Journal of biological chemistry
Volume263
Issue number33
StatePublished - Jan 1 1988
Externally publishedYes

Fingerprint

Phosphoenolpyruvate
Nucleic Acid Regulatory Sequences
Retroviridae
Genetic Promoter Regions
Hepatocellular Carcinoma
Genes
Glucocorticoids
Terminal Repeat Sequences
Proviruses
Phosphoenolpyruvate Carboxykinase (GTP)
Tyrosine Transaminase
Cells
Regulator Genes
Population
Phosphotransferases
Genome
Hormones
Rats
RNA
Insulin

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hormonal regulation of chimeric genes containing the phosphoenolpyruvate carboxykinase promoter regulatory region in hepatoma cells infected by murine retroviruses. / Hatzoglou, M.; Park, Edwards; Wynshaw-Boris, A.; Kaung, H. L.; Hanson, R. W.

In: The Journal of biological chemistry, Vol. 263, No. 33, 01.01.1988, p. 17798-17808.

Research output: Contribution to journalArticle

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