Abstract
The effect of transient transfection with expression vectors of Ha-ras on the promoter activity of 12-lipoxygenase in human epidermoid carcinoma A431 cells was studied. Overexpression of Ha-ras increased the promoter activity in a dose- and time-dependent manner, which correlated closely with the cellular expression of Ras protein. Promoters of different gene lengths for human 12-lipoxygenase were used to prepare the luciferase fusion vectors. Following transfection by Ha-ras for 68 h, an approx. 40-fold increase in luciferase reporter activity was observed in plasmids with the 5'-flanking region ranging from -951 to -224 bp upstream from translation starting site. There was no obvious stimulation in cells transfected with a vector-bearing promoter with a length of -100 bp. These results indicate that the promoter region ranging from -224 to -100 bp was important for the Ha-ras response. With the aid of additional 5'-deletion and site-directed mutagenesis, three Sp1 binding sequences residing at -169 to -161 bp, -158 to -150 bp and -123 to -114 bp were found to be critical for the Ha-ras response of activating the transcription of human 12-lipoxygenase gene.
| Original language | English |
|---|---|
| Pages (from-to) | 270-277 |
| Number of pages | 8 |
| Journal | Biochimica et Biophysica Acta - Lipids and Lipid Metabolism |
| Volume | 1344 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Feb 18 1997 |
| Externally published | Yes |
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Keywords
- 12-Lipoxygenase
- Human epidermoid carcinoma A431 cell
- Ras protein
- Transcription
ASJC Scopus subject areas
- Biochemistry
- Endocrinology
- Biophysics
Cite this
Overexpression of Ha-ras enhances the transcription of human arachidonate 12-lipoxygenase promoter in A431 cells. / Chen, Ben Kuen; Liu, Yi Wen; Yamamoto, Shozo; Chang, Wen Chang.
In: Biochimica et Biophysica Acta - Lipids and Lipid Metabolism, Vol. 1344, No. 3, 18.02.1997, p. 270-277.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Overexpression of Ha-ras enhances the transcription of human arachidonate 12-lipoxygenase promoter in A431 cells
AU - Chen, Ben Kuen
AU - Liu, Yi Wen
AU - Yamamoto, Shozo
AU - Chang, Wen Chang
PY - 1997/2/18
Y1 - 1997/2/18
N2 - The effect of transient transfection with expression vectors of Ha-ras on the promoter activity of 12-lipoxygenase in human epidermoid carcinoma A431 cells was studied. Overexpression of Ha-ras increased the promoter activity in a dose- and time-dependent manner, which correlated closely with the cellular expression of Ras protein. Promoters of different gene lengths for human 12-lipoxygenase were used to prepare the luciferase fusion vectors. Following transfection by Ha-ras for 68 h, an approx. 40-fold increase in luciferase reporter activity was observed in plasmids with the 5'-flanking region ranging from -951 to -224 bp upstream from translation starting site. There was no obvious stimulation in cells transfected with a vector-bearing promoter with a length of -100 bp. These results indicate that the promoter region ranging from -224 to -100 bp was important for the Ha-ras response. With the aid of additional 5'-deletion and site-directed mutagenesis, three Sp1 binding sequences residing at -169 to -161 bp, -158 to -150 bp and -123 to -114 bp were found to be critical for the Ha-ras response of activating the transcription of human 12-lipoxygenase gene.
AB - The effect of transient transfection with expression vectors of Ha-ras on the promoter activity of 12-lipoxygenase in human epidermoid carcinoma A431 cells was studied. Overexpression of Ha-ras increased the promoter activity in a dose- and time-dependent manner, which correlated closely with the cellular expression of Ras protein. Promoters of different gene lengths for human 12-lipoxygenase were used to prepare the luciferase fusion vectors. Following transfection by Ha-ras for 68 h, an approx. 40-fold increase in luciferase reporter activity was observed in plasmids with the 5'-flanking region ranging from -951 to -224 bp upstream from translation starting site. There was no obvious stimulation in cells transfected with a vector-bearing promoter with a length of -100 bp. These results indicate that the promoter region ranging from -224 to -100 bp was important for the Ha-ras response. With the aid of additional 5'-deletion and site-directed mutagenesis, three Sp1 binding sequences residing at -169 to -161 bp, -158 to -150 bp and -123 to -114 bp were found to be critical for the Ha-ras response of activating the transcription of human 12-lipoxygenase gene.
KW - 12-Lipoxygenase
KW - Human epidermoid carcinoma A431 cell
KW - Ras protein
KW - Transcription
UR - http://www.scopus.com/inward/record.url?scp=0031576833&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031576833&partnerID=8YFLogxK
U2 - 10.1016/S0005-2760(96)00151-8
DO - 10.1016/S0005-2760(96)00151-8
M3 - Article
VL - 1344
SP - 270
EP - 277
JO - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
JF - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
SN - 1388-1981
IS - 3
ER -