Identification of a functional transcriptional factor AP-1 site in the sheep interferon τ gene that mediates a response to PMA in JEG3 cells

Hirohito Yamaguchi, Yasuhiro H Ikeda, J. Ignacio Moreno, Momoko Katsumura, Takayuki Miyazawa, Eiji Takahashi, Kazuhiko Imakawa, Senkiti Sakai, Ronald K. Christenson

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

To examine regulatory mechanisms of sheep interferon τ (oIFNτ) gene expression, potential enhancer/silencer elements of the oIFNτ gene were examined using a transient transfection system with oIFNτ gene-chloramphenicol acetyltransferase (oIFNτ-CAT) reporter constructs in human choriocarcinoma cells, JEG3. Experiments with 5'-deletion constructs revealed that the upstream regions from bases -654 to -607 and from bases -606 to -555 were essential for oIFNτ gene expression. In a heterologous transcriptional system in which the upstream regions of oIFNτ were inserted in front of simian virus 40 (SV40) promoter, the regions between bases -654 and -555 were determined as being the enhancer region required for oIFNτ-SV40-CAT transactivation. A subsequent study with the oIFNτ-CAT constructs lacking the upstream region between bases -542 and -124 revealed that, in addition to the further upstream region between bases -1000 and -654, the sequences from bases -543 to -452 seemed to act as silencer regions. The oIFNτ-CAT constructs with site-specific mutagenesis revealed that multiple enhancer elements existed between bases -654 and -555 of the oIFNτ gene. On the basis of nucleotide sequence analysis, there are numerous sites between bases -654 and -555 to which potential transcriptional factors, AP-1, GATA and GATA-related proteins, could bind. Furthermore, gel mobility-shift assays revealed that AP-1 or other nuclear factors could bind to these elements. In co-transfection studies, the expression of c-Jun plus c-Fos enhanced the transactivation of oIFNτ-CAT but the expression of GATA-1, GATA-2 or GATA-3 did not. Taken together, these results suggest that the upstream region between bases -654 and -555 could be considered as the enhancer region for oIFNτ gene transactivation.

Original languageEnglish (US)
Pages (from-to)767-773
Number of pages7
JournalBiochemical Journal
Volume340
Issue number3
DOIs
StatePublished - Jun 15 1999
Externally publishedYes

Fingerprint

Transcription Factor AP-1
Interferons
Sheep
Genes
Transcriptional Activation
Simian virus 40
Viruses
Gene expression
Transfection
Transcriptional Silencer Elements
Gene Expression
Mutagenesis
Choriocarcinoma
Chloramphenicol O-Acetyltransferase
Electrophoretic Mobility Shift Assay
Site-Directed Mutagenesis
Genetic Promoter Regions
Sequence Analysis
Assays
Nucleotides

Keywords

  • CAT reporter gene
  • Enhancer elements
  • Interferon gene expression
  • Transcription

ASJC Scopus subject areas

  • Biochemistry

Cite this

Yamaguchi, H., Ikeda, Y. H., Moreno, J. I., Katsumura, M., Miyazawa, T., Takahashi, E., ... Christenson, R. K. (1999). Identification of a functional transcriptional factor AP-1 site in the sheep interferon τ gene that mediates a response to PMA in JEG3 cells. Biochemical Journal, 340(3), 767-773. https://doi.org/10.1042/0264-6021:3400767

Identification of a functional transcriptional factor AP-1 site in the sheep interferon τ gene that mediates a response to PMA in JEG3 cells. / Yamaguchi, Hirohito; Ikeda, Yasuhiro H; Moreno, J. Ignacio; Katsumura, Momoko; Miyazawa, Takayuki; Takahashi, Eiji; Imakawa, Kazuhiko; Sakai, Senkiti; Christenson, Ronald K.

In: Biochemical Journal, Vol. 340, No. 3, 15.06.1999, p. 767-773.

Research output: Contribution to journalArticle

Yamaguchi, H, Ikeda, YH, Moreno, JI, Katsumura, M, Miyazawa, T, Takahashi, E, Imakawa, K, Sakai, S & Christenson, RK 1999, 'Identification of a functional transcriptional factor AP-1 site in the sheep interferon τ gene that mediates a response to PMA in JEG3 cells', Biochemical Journal, vol. 340, no. 3, pp. 767-773. https://doi.org/10.1042/0264-6021:3400767
Yamaguchi, Hirohito ; Ikeda, Yasuhiro H ; Moreno, J. Ignacio ; Katsumura, Momoko ; Miyazawa, Takayuki ; Takahashi, Eiji ; Imakawa, Kazuhiko ; Sakai, Senkiti ; Christenson, Ronald K. / Identification of a functional transcriptional factor AP-1 site in the sheep interferon τ gene that mediates a response to PMA in JEG3 cells. In: Biochemical Journal. 1999 ; Vol. 340, No. 3. pp. 767-773.
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