Activity of hepatocyte nuclear factor l alpha and hepatocyte nuclear factor 1 beta isoforms is differently affected by the inhibition of protein phosphatases 1/2A

Phosphorylation/dephosphorylation processes are known to control the activi ty of several transcription factors. The nutrition-dependent expression of sucrase-isomaltase and Na+/glucose cotransporter 1, two proteins implicated in the intestinal absorption of glucose, has been shown to be closely rela ted to modifications of hepatocyte nuclear factor 1 (HNF1) activity. This s tudy was conducted to determine whether phosphorylation/dephosphorylation p rocesses could control HNF1 activity. We show that expression of the gene e ncoding sucrase-isomaltase is inhibited in the enterocytic Caco-2 clone TC7 by okadaic acid at a concentration that is known to inhibit protein phosph atases 1/2A and that does not affect cell viability. At the same concentrat ion, phosphorylation of the HNF1 alpha and HNF1 beta isoforms is greatly en hanced and their DNA-binding capacity is decreased. The phosphorylation sta te of HNF1 beta isoforms directly affects their DNA-binding capacity. In co ntrast, the decreased DNA-binding activity of the HNF1 alpha isoforms, whic h was observed after the inhibition of protein phosphatases 1/2A, is due to a net decrease in their total cellular and nuclear amounts. Such an effect results from a decrease in both the HNF1 alpha mRNA levels and the half-li fe of the protein. This is the first evidence for the implication of protei n phosphatases 1/2A in the control of the activity of HNF1 isoforms. Moreov er, these results emphasize a physiological role for the balance between ph osphatases and kinases in the nutrition-dependent regulation of HNF1-contro lled genes.