Metal transcription factor-1 regulation via MREs in the transcribed regions of selenoprotein H and other metal-responsive genes.
Selenoprotein H is a redox-sensing DNA binding protein that upregulates genes involved in antioxidant responses. Given the known links between oxidative stress and heavy metals, we investigated the potential for regulation of selenoprotein H by metals. In silico analysis of the selenoprotein H genes from nine species reveals multiple predicted metal response elements (MREs). To validate MRE function, we investigated the effects of zinc or cadmium addition and metal-responsive transcription factor 1 (MTF-1) knockout on selenoprotein H mRNA levels. Chromatin immunoprecipitation was used to directly assess physical binding of the transcription factor to MREs in the human and mouse selenoprotein H genes. The results reported herein show that selenoprotein H is a newly identified target for MTF-1. Further, whereas nearly all prior studies of MREs focused on those located in promoters, we demonstrate binding of MTF-1 to MREs located downstream of the transcription start sites in the human and murine selenoprotein H genes. Finally, we identified MREs in downstream sequences in 15 additional MTF-1 regulated genes lacking promoter MREs, and demonstrated MTF-1 binding in three of these genes. This regulation via sequences downstream of promoters highlights a new direction for identifying previously unrecognized target genes for MTF-1.
Stoytcheva
Zoia R
ZR
Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii at Manoa, 651 Ilalo Street, Suite 222, Honolulu, HI 96813, USA.
Vladimirov
Vladimir
V
Douet
Vanessa
V
Stoychev
Ilko
I
Berry
Marla J
MJ
D000650:Amnion / Q000502:physiology; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D017930:Genes, Reporter; D006801:Humans; D007668:Kidney; D008156:Luciferases / Q000235:genetics; D008251:Macaca; D019216:Metals, Heavy / Q000633:toxicity; D051379:Mice; D018384:Oxidative Stress; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051140:Selenoproteins / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D015854:Up-Regulation; D000097809:Transcription Factor MTF-1
D017931:DNA Primers; D004268:DNA-Binding Proteins; D008156:Luciferases; D019216:Metals, Heavy; D051140:Selenoproteins; D014157:Transcription Factors; D000097809:Transcription Factor MTF-1; C519118:selenoprotein H, human; C547924:selenoprotein H, mouse