PU.1, interferon regulatory factor 1, and interferon consensus sequence-binding protein cooperate to increase gp91(phox) expression.
gp91(phox) is a subunit of the phagocyte respiratory burst oxidase catalytic unit. Transcription of CYBB, the gene encoding gp91(phox), is restricted to terminally differentiated phagocytic cells. An element in the proximal CYBB promoter binds a protein complex, referred to as hematopoiesis-associated factor (HAF1), that is necessary for interferon-gamma (IFNgamma)-induced gp91(phox) expression. In these investigations, we determined that HAF1 was a multiprotein complex, cross-immunoreactive with the transcription factors PU.1, interferon regulatory factor 1 (IRF-1), and interferon consensus sequence-binding protein (ICSBP). In electrophoretic mobility shift assay, the HAF1 complex was reconstituted by either in vitro translated PU.1 with IRF-1 or PU.1 with ICSBP, but not by IRF-1 with ICSBP. HAF1a, a slower mobility complex with the same binding site specificity as HAF1, was also investigated. Similar to the HAF1 complex, the HAF1a complex was cross-immunoreactive with PU. 1, IRF-1, and ICSBP. Unlike the HAF1 complex, reconstitution of the HAF1a complex required in vitro translated PU.1 with both IRF-1 and ICSBP. An artificial promoter construct containing the HAF1/HAF1a binding site was modestly activated in the myelomonocytic cell line U937 by co-transfection either with PU.1 and IRF-1 or with PU.1 and ICSBP, but it was strongly activated by co-transfection with PU.1, IRF-1, and ICSBP. This activation required serine 148-phosphorylated PU.1. These studies describe a novel mechanism for PU.1 transcriptional activation via interaction with both IRF-1 and ICSBP, a target gene for the interaction of IRF-1 with ICSBP, and a novel activation function for ICSBP as a component of a multiprotein complex.
Eklund
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EA
Lurleen B. Wallace Tumor Institute, Department of Hematology and Oncology and the Comprehensive Cancer Center, University of Alabama at Birmingham and the Birmingham Veterans Administration Hospital, Birmingham, Alabama 35294, USA.
Jalava
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A
Kakar
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R
D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D050856:Interferon Regulatory Factor-1; D050835:Interferon Regulatory Factors; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D010750:Phosphoproteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D015534:Trans-Activators / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D000099185:Interferon Regulatory Factor-8; D000099232:Proto-Oncogene Protein Spi-1
D004268:DNA-Binding Proteins; D050856:Interferon Regulatory Factor-1; D050835:Interferon Regulatory Factors; D008562:Membrane Glycoproteins; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D010750:Phosphoproteins; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D000099185:Interferon Regulatory Factor-8; D000099232:Proto-Oncogene Protein Spi-1; C494257:IRF1 protein, human; C056558:CYBB protein, human