Adverse effect of cadmium on binding of transcription factor Sp1 to the GC-rich regions of the mouse sodium-glucose cotransporter 1, SGLT1, promoter.
Exposure of the kidney to cadmium can cause glucosuria. Effect of cadmium on sodium-glucose cotransporter 1, (SGLT1) mRNA molecules in cultured mouse kidney cortical cells was determined by quantitative competitive RT-PCR. SGLT1 mRNA molecules decreased from 58 x 10(4) microg(-1) total RNA in untreated cells to 29 x 10(4) microg(-1) total RNA in cells exposed to 5 microM cadmium. Increasing cadmium to 7.5 and 10 microM, reduced mRNA molecules to 21 x 10(4) and 12 x 10(4) microg(-1) total RNA, respectively. The half-life of SGLT1 mRNA in control and in cells exposed to 7.5 microM cadmium were almost the same and calculated to be 9.1 h (S.E.+/-2.7) for the former and 8.5 h (S.E.+/-2.2) for the latter. We also analyzed mouse SGLT1 promoter sequences and identified two conserved Sp1 binding sites. The Sp1 binding sequences were used as probes in electrophoretic mobility shift assay (EMSA) with nuclear proteins from cultured cells. Intensity of complexes of the 5' and the 3' Sp1 probes with nuclear Sp1 from cells treated with 7.5 microM cadmium were 84% (S.E.+/-4) and 61% (S.E.+/-14) of controls, respectively. Cadmium had no effect on expression of Sp1 mRNA or protein level. Cadmium-induced inhibition of glucose uptake in kidney may be the result of transcriptional down-regulation of SGLT1 mediated through modification of Sp1 binding to its promoter.
Tabatabai
Niloofar M
NM
Department of Chemistry, University of Wisconsin-Milwaukee, PO Box 413, Milwaukee, WI 53201, USA. niloofar@uwm.edu
Blumenthal
Samuel S
SS
Petering
David H
DH
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002104:Cadmium / Q000633:toxicity*; D002105:Cadmium Poisoning / Q000209:etiology / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D006801:Humans; D007672:Kidney Cortex / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D051269:Sodium-Glucose Transporter 1; D016329:Sp1 Transcription Factor / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
D008562:Membrane Glycoproteins; D009004:Monosaccharide Transport Proteins; D012333:RNA, Messenger; C495674:SLC5A1 protein, human; C495675:Slc5a1 protein, mouse; D051269:Sodium-Glucose Transporter 1; D016329:Sp1 Transcription Factor; D002104:Cadmium