PU.1 regulates the expression of the human neutrophil elastase gene.
PU.1 is a transcription factor present in B-cells and macrophages. Here, we report our studies on the role of PU.1 in myelopoiesis using human neutrophil elastase (HNE) as a model. HNE, a component of the primary granules of mature granulocytes, is a serine protease which is transcriptionally restricted to the late promyelocytic stage of granulocytic maturation. The first 200 bp of the HNE promoter directs myeloid specific expression of a reporter gene and a 30-bp element within this region was been identified as the major determinant of myeloid specific expression [S. Srikanth, T. Rado, A 30-bp element is responsible for the myeloid specific activity of the human neutrophil elastase promoter, J. Biol. Chem. 269 (1994) 32626-32632.]. We now show that the B-cell and macrophage specific transcription factor, PU.1, binds to the PU.1 consensus site within the 30-bp element to activate transcription. Substitution mutations within this recognition sequence results in the loss of PU.1 binding and in a 90% decrease in promoter activity in myeloid cells. Cotransfection of PU.1 and a reporter gene controlled by the HNE promoter into non-myeloid HeLa cells resulted in activation of reporter gene transcription.
Department of Medicine, University of Colorado Health Sciences Center, Denver 80262, USA.
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D019943:Amino Acid Substitution; D000906:Antibodies / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D002457:Cell Extracts / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D005796:Genes / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D019272:Leukocyte Elastase / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D015534:Trans-Activators / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D014162:Transfection / Q000235:genetics; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism; D000099232:Proto-Oncogene Protein Spi-1
D000906:Antibodies; D002457:Cell Extracts; D004247:DNA; D004268:DNA-Binding Proteins; D019272:Leukocyte Elastase; D010744:Phosphoric Monoester Hydrolases; D011518:Proto-Oncogene Proteins; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1