The 5'region of the rat phosphoenolpyruvate carboxykinase gene confers pH sensitivity to chimeric genes expressed in renal and liver cell lines capable of expressing PEPCK.
The 5' flanking regions of the rat phosphoenolpyruvate carboxykinase gene were used to form chimeric gene constructs with the human growth hormone gene. These constructs were transfected into several renal and one liver cell line and the production of growth hormone (HGH) measured by immunoassay. Cyclic-AMP and glucocorticoid responsiveness of HGH production was observed in all cell lines. In two lines, the rat NRK52E renal epithelial line and the rat H4IIE hepatoma cell line, both capable of expressing PEPCK, lowering extracellular pH increased HGH production several fold. Comparison of hormone and pH effect on cells transfected with a thymidine kinase promoter-HGH chimera indicated that the PEPCK 5' flanking region effects were specific. Thus, part of the pH responsiveness of the PEPCK gene in vivo may be attributed to properties of the 5' flanking regions.
Pollock
A S
AS
Department of Medicine/Nephrology, University of California, San Francisco.
Long
J A
JA
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D000242:Cyclic AMP / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology; D015870:Gene Expression; D005938:Glucocorticoids / Q000494:pharmacology; D013006:Growth Hormone / Q000096:biosynthesis / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000166:cytology / Q000201:enzymology*; D008099:Liver / Q000166:cytology / Q000201:enzymology*; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000235:genetics*; D051381:Rats; D014162:Transfection
D005938:Glucocorticoids; D013006:Growth Hormone; D000242:Cyclic AMP; D010729:Phosphoenolpyruvate Carboxykinase (GTP)