Identification and characterization of a novel hypertonicity-responsive element in the human aquaporin-1 gene

The AQP1 gene is transcriptionally upregulated in response to hypertonicity. However, the molecular mechanism in hypertonicity-induced transcription in the AQP1 gene has been poorly understood. Here we report a novel hypertonicity response element (HRE) in the AQP1 gene. A critical cis-acting element to hypertonicity in the AQP1 promoter is located at -54 to -46 and the change from GCTCCCCCC to GCTTTCCCC completely abolished the hypertonic induction and osmotic response, thus implicating the importance of the first CC sequence in the region. Two prominent DNA-protein complexes were observed electrophoretic mobility shift assay (EMSA), and the band intensities in nuclear extracts from osmotically stressed cells were much higher than that of isotonic nuclear extracts. The EMSA supershift assay with anti-Sp1 antibody showed that two retarded bands did not bind to anti-Sp1 antibody, suggesting that the DNA binding proteins bound to the HRE are not Sp1 family proteins. These data suggest that the transcription of the AQP1 by hypertonicity in renal cells is upregulated by the interaction with putative DNA binding proteins to a novel HRE located at -54 to -46 in the AQP1 gene. (C) 2002 Elsevier Science (USA).