Control of placental alkaline phosphatase gene expression in HeLa cells: induction of synthesis by prednisolone and sodium butyrate.
HeLa S3 cells produce an alkaline phosphatase indistinguishable from the enzyme from human term placenta. The phosphatase activity in these cells was induced by both prednisolone and sodium butyrate. Both agents stimulated de novo synthesis of the enzyme. The increase in phosphatase activity paralleled the increase in immunoactivity and biosynthesis of placental alkaline phosphatase. The fully processed phosphatase monomer in control, prednisolone-treated or butyrate-treated cells was a 64.5 K polypeptide, measured by both incorporation of L-[35S]methionine into enzyme protein and active-site labeling. The 64.5 K polypeptide was formed by the incorporation of additional N-acetylneuraminic acid moieties to a precursor polypeptide of 61.5 K. However, this biosynthetic pathway was identified only in butyrate-treated cells. In prednisolone-treated cells, the processing of 61.5 K to the 64.5 K monomer was accelerated, and the presence of the 61.5 K precursor could only be detected by either neuraminidase or monensin treatment. Phosphatase mRNA which comigrated with the term placental alkaline phosphatase mRNA of 2.7 kilobases was induced in the presence of either prednisolone or butyrate. Alkaline phosphatase mRNA in untreated HeLa S3 cells migrated slightly faster than the term placental alkaline phosphatase mRNA. Butyrate also induced a second still faster migrating alkaline phosphatase mRNA. Both prednisolone and butyrate increased the steady-state levels of placental alkaline phosphatase mRNA. Our data indicate that the increase in phosphatase mRNA by prednisolone and butyrate resulted in the induction of alkaline phosphatase activity and biosynthesis in HeLa S3 cells. Furthermore, both agents induced the expression of different alkaline phosphatase gene transcripts without altering its protein product.
D000469:Alkaline Phosphatase / Q000096:biosynthesis* / Q000235:genetics; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D004247:DNA / Q000032:analysis; D004790:Enzyme Induction; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D005796:Genes / Q000187:drug effects*; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D009693:Nucleic Acid Hybridization; D010920:Placenta / Q000201:enzymology*; D011239:Prednisolone / Q000494:pharmacology*
D002087:Butyrates; D007527:Isoenzymes; D020148:Butyric Acid; D004247:DNA; D011239:Prednisolone; D000469:Alkaline Phosphatase