title: Cadmium down-regulation of kidney Sp1 binding to mouse SGLT1 and SGLT2 gene promoters: possible reaction of cadmium with the zinc finger domain of Sp1.
abstract: Cadmium (Cd) exposure causes glucosuria (glucose in the urine). Previously, it was shown that Cd exposure of primary cultures of mouse kidney cells (PMKC) decreased mRNA levels of the glucose transporters, SGLT1 and SGLT2 and that Sp1 from Cd-exposed cells displayed reduced binding to the GC boxes of the mouse SGLT1 promoter in vitro. Here, we identified a GC box upstream of mouse SGLT2 gene. ChIP assays on PMKC revealed that exposure to 5 microM Cd abolished Sp1 binding to SGLT1 GC box while it decreased Sp1 binding to SGLT2 GC sequence by 30% in vivo. The in vitro DNA binding assay, EMSA, demonstrated that binding of Sp1 from Cd (7.5 microM)-treated PMKC to the SGLT2 GC probe was 86% lower than in untreated cells. Sp1 is a zinc finger protein. Compared to PMKC exposed to 5 microM Cd alone, inclusion of 5 microM Zn restored SGLT1 and 2 mRNA levels by 15% and 30%, respectively. Cd (10 microM) decreased the binding of recombinant Sp1 (rhSp1) to SGLT1 and SGLT2 GC probes to 12% and 8% of untreated controls. Cd exerted no effect on GC-bound rhSp1. Co-treatment with Cd and Zn showed that added Zn significantly restored rhSp1 binding to the SGLT1 and SGLT2. Addition of Zn post Cd treatment was not stimulatory. We conclude that Cd can replace Zn in Sp1 DNA binding domain to reduce its binding to GC sites in mouse SGLT1 and SGLT2 promoters.
authors.affiliation: Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee, Milwaukee, WI 53201, USA.
authors.forename: Rajendra K
authors.initials: RK
authors.lastname: Kothinti
authors.forename: Amy B
authors.initials: AB
authors.lastname: Blodgett
authors.forename: David H
authors.initials: DH
authors.lastname: Petering
authors.forename: Niloofar M
authors.initials: NM
authors.lastname: Tabatabai
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000187:drug effects; D002104:Cadmium / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics; D004785:Environmental Pollutants / Q000378:metabolism / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D051269:Sodium-Glucose Transporter 1 / Q000235:genetics* / Q000378:metabolism; D051297:Sodium-Glucose Transporter 2 / Q000235:genetics* / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000378:metabolism; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers / Q000187:drug effects
chemical_list: D004785:Environmental Pollutants; C495675:Slc5a1 protein, mouse; C490299:Slc5a2 protein, mouse; D051269:Sodium-Glucose Transporter 1; D051297:Sodium-Glucose Transporter 2; D016329:Sp1 Transcription Factor; D002104:Cadmium; D015032:Zinc