Glucose regulates the promoter activity of aldolase A and pyruvate kinase M2 via dephosphorylation of Sp1.
Proliferating cells and tumour cells maintain a high glycolytic rate even under aerobic conditions. FTO2B cells, a rat hepatoma cell line, show high activities of glycolytic enzymes. Within a culture period of 48 h the cell number increases 5-fold. Replacement of glucose by pyruvate in the culture medium lowers glycolytic enzyme activity and prevents proliferation. Transfection assays revealed that glucose deprivation dramatically decreases the transcriptional activities of the Sp1-dependent aldolase and pyruvate kinase promoters leading to reduced reporter gene expression. Sp1 binding activity is also inhibited by ocadaic acid, an inhibitor of protein phosphatase 1. Western blot analyses with nuclear extracts from FTO2B cells cultured in the presence or absence of glucose revealed differences in the phosphorylation state of Sp1. From these results we conclude that glucose increases the amount of the dephosphorylated form of Sp1 which has a higher DNA binding activity. As a consequence gene expression of the glycolytic enzymes is increased which is a prerequisite for cell proliferation.
Schäfer
D
D
Institut für Biochemie, Medizinische Fakultät, Universität Erlangen-Nürnberg, Erlangen, Germany.
Hamm-Künzelmann
B
B
Brand
K
K
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D005634:Fructose-Bisphosphate Aldolase / Q000096:biosynthesis* / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology*; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D011770:Pyruvate Kinase / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D007527:Isoenzymes; D011994:Recombinant Proteins; D016329:Sp1 Transcription Factor; D011770:Pyruvate Kinase; D005634:Fructose-Bisphosphate Aldolase; D005947:Glucose