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Differential effect of epidermal growth factor on mitochondrial glycerophospate acyltransferase distal and proximal promoters
Author(s) -
Guha Prajna,
Aneja Kawalpreet K,
Gaddi Pamela,
Chakraborty Sanjoy,
Haldar Dipak
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.lb44-c
Subject(s) - luciferase , transfection , promoter , microbiology and biotechnology , epidermal growth factor , reporter gene , chemistry , biology , cell culture , gene , gene expression , biochemistry , genetics
We have previously cloned the proximal and distal promoters of mitochondrial glycerophosphate acyltransferase (mt‐GPAT) in pGL3 basic vector containing luciferase as the reporter gene. When COS‐1 cells were transfected with these constructs and starved and refed with glucose, the luciferase activity was 6 to 8 times more in cells containing the distal but not the proximal promoter. In the present investigation, the effect of epidermal growth factor (EGF) on the mt‐GPAT distal and proximal promoters was examined. Rat liver epithelial cells were transfected with the cloned pGL3 constructs containing either the distal or the proximal promoter and incubated in the absence and presence of EGF. The cell lysates were analyzed for luciferase activity. Two experimental parameters were tested – hours of incubation and concentration of EGF. The maximum luciferase activity, over 3 times of control, was observed when the cells transfected with pGL3 vector with the distal promoter were incubated for 24 hours with EGF at a concentration of 50ng/ml. There was no significant change in the luciferase activity for cells transfected with pGL3 vector containing the proximal promoter under any of the above conditions. These results provide evidence that the distal, and not the proximal promoter, is inducible by EGF. Supported by NIH grant GM‐57643 and Estee Lauder.