Dual regulation of ets-activated gene expression by SP1.
The human folate receptor (hFR) type gamma gene is driven by a TATA-less promoter that uses a canonical Sp1 element for basal transcription. Using nuclear extract from 293 (human embryonic) cells, we mapped a second (non-canonical) Sp1 element to which Sp1 bound with a comparable affinity and which overlaps a functional ets binding site (EBS). Mutagenesis experiments revealed that the binding of ets to the EBS activates the promoter synergistically with Sp1 bound at the downstream site; however, binding of Sp1 to the EBS does not contribute to promoter activity. A further increase in Sp1 by inducible expression in recombinant 293 cells resulted in a small but significant decrease in the hFR-gamma promoter activity, but the decrease was abolished when the EBS was deleted from the promoter. In 293 cells, which do not express hFR-gamma, the Sp1 level was relatively high whereas in the hFR-gamma-positive HL60 leukemia cells, the Sp1 level was low and the EBS predominantly bound an ets protein. To account for the above observations, we propose a model in which when the Sp1 level is low, ets out competes Sp1 for binding to the EBS and synergistically enhances the hFR-gamma promoter activity by interacting with Sp1 bound at the canonical site whereas at higher levels, Sp1 represses the promoter by competitively inhibiting the binding of ets. As a partial extension of this model to the regulation of other ets activated genes, we show that Sp1 can predictably bind to a variety of ets elements including those responsive to Ets1 and Spi.1/Pu.1. A dual concentration-dependent action of Sp1 as an activator or a repressor offers a potential mechanism contributing to tissue-specific regulation of ets-dependent genes by Sp1.
Kelley
Karen M M
KM
Department of Biochemistry and Molecular Biology, Medical College of Ohio, 3035 Arlington Avenue, Toledo, OH 43614-5804, USA.
Wang
Huiquan
H
Ratnam
Manohar
M
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D024202:Electrophoretic Mobility Shift Assay; D058973:Folate Receptors, GPI-Anchored; D005786:Gene Expression Regulation / Q000235:genetics*; D018922:HL-60 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009154:Mutation; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D050783:Proto-Oncogene Proteins c-ets; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
D002352:Carrier Proteins; C494041:ETS1 protein, human; C494042:Ets1 protein, mouse; D058973:Folate Receptors, GPI-Anchored; D009841:Oligonucleotides; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; D008156:Luciferases