Hormonal control of interacting promoters introduced into cells by retroviruses

Maria Hatzoglou, Fatima Bosch, Edwards Park, Richard W. Hanson

Research output: Contribution to journalArticle

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Abstract

The interaction of promoters contained in a Moloney murine leukemia virus (MoMLV)-based retroviral vector was studied after infection of FTO-2B rat hepatoma and NIH 3T3 mouse fibroblast cells. Segments of the phosphoenolpyruvate carboxykinase (PEPCK) promoter-regulatory region, which are known from previous studies to confer responsiveness to hormones, were linked to the structural genes for bovine growth hormone, amino-3′-glycosylphosphotransferase (neo), and herpes-virus thymidine kinase and inserted into a MoMLV-based retroviral vector. In vectors in which PEPCK was the only internal promoter, it was the major site of gene transcription. This dominant effect was independent of the orientation of the PEPCK promoter relative to the 5′ long terminal repeat of the provirus and was noted with as little as -174 base pairs of the 5′-flanking sequence. NIH 3T3 cells, which do not express the endogenous PEPCK gene, transcribed the transduced PEPCK-chimeric genes at the same high levels as was observed in hepatoma cells. When two promoters were present in the provirus, the expression of chimeric structural genes depended on the relative position and orientation of these genes as well as the type of cell infected by the retrovirus. Differential responses of proviral promoters in infected cells were also observed in the presence of hormones. Dibutyryl cyclic AMP increased the expression of genes linked to the PEPCK promoter in FTO-2B and NIH 3T3 cells, whereas glucocorticoids stimulated transcription from both the PEPCK promoter and the long terminal repeat in FTO-2B cells. The effect of these hormones on transcription of proviral promoters depended on their position relative to the 5′ long terminal repeat. In contrast, insulin uniformly inhibited transcription from the PEPCK promoter in a position-independent manner but only in hepatoma cells and not in fibroblasts. In clonally isolated FTO-2B cells infected with a retrovirus, the site of proviral integration was also a major factor determining the expression and hormonal regulation from the internal promoters. The data suggest that the hormonal regulation of the expression of genes contained in retroviral vectors depends on the type and position of the regulatory elements present in the provirus and the lineage of the infected cell.

Original languageEnglish (US)
Pages (from-to)8416-8425
Number of pages10
JournalJournal of Biological Chemistry
Volume266
Issue number13
StatePublished - May 5 1991
Externally publishedYes

Fingerprint

Phosphoenolpyruvate
Retroviridae
Genes
Transcription
Proviruses
Terminal Repeat Sequences
Viruses
Moloney murine leukemia virus
Hepatocellular Carcinoma
NIH 3T3 Cells
Hormones
Fibroblasts
Bucladesine
Gene Order
Thymidine Kinase
5' Flanking Region
Nucleic Acid Regulatory Sequences
Gene Expression Regulation
Genetic Promoter Regions
Base Pairing

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Hormonal control of interacting promoters introduced into cells by retroviruses. / Hatzoglou, Maria; Bosch, Fatima; Park, Edwards; Hanson, Richard W.

In: Journal of Biological Chemistry, Vol. 266, No. 13, 05.05.1991, p. 8416-8425.

Research output: Contribution to journalArticle

Hatzoglou, M, Bosch, F, Park, E & Hanson, RW 1991, 'Hormonal control of interacting promoters introduced into cells by retroviruses', Journal of Biological Chemistry, vol. 266, no. 13, pp. 8416-8425.
Hatzoglou, Maria ; Bosch, Fatima ; Park, Edwards ; Hanson, Richard W. / Hormonal control of interacting promoters introduced into cells by retroviruses. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 13. pp. 8416-8425.
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