NF-Y, a CCAAT box-binding protein, is one of the trans-acting factors necessary for the response of the murine ERp72 gene to protein traffic.
The accumulation of incompletely assembled immunoglobulin mu heavy chain in transfected COS cells stimulates the cellular response to protein traffic that results in the increased transcription and elevated synthesis of several ER chaperones, including ERP72, a member of the protein disulfide isomerase family of molecular chaperones. The ERp72 promoter contains an 82 bp ER protein traffic response element (ERPTRE) that is sufficient to mediate this response. Previously, it had been shown that the alteration of a putative AP-2 site and a CCAAT and inverted CCAAT site within the ERPTRE significantly decreased the response of ERp72 promoter to mu chain accumulation. We have extended these findings by demonstrating a role for NF-Y and a potentially novel DNA-binding protein in the regulation of transcription from the ERp72 promoter. The fact that NF-Y binding to the ERPTRE is observed in extracts from both control cells and cells in which the response to protein traffic has been activated indicates that the binding of NF-Y, while necessary, is not sufficient to account for the response. Each of the two CCAAT sites in the ERPTRE can bind NF-Y independently, but both sites must be intact for full ERPTRE function. A second protein can bind to the ERPTRE independently of NF-Y and at a site overlapping or close to the 3' end of the reverse CCAAT site. It is possible that interactions between NF-Y, this protein and perhaps other factors are responsible for the regulation of the protein traffic response.
Marcus
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N
Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, MO 63104, USA.
Green
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M
D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D022762:CCAAT-Enhancer-Binding Proteins; D019556:COS Cells; D000001:Calcimycin / Q000494:pharmacology; D002457:Cell Extracts; D002467:Cell Nucleus; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D007148:Immunoglobulin mu-Chains / Q000737:chemistry / Q000378:metabolism; D007476:Ionophores / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D018832:Molecular Chaperones / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D017510:Protein Folding; D014157:Transcription Factors / Q000378:metabolism*; D014466:Ultraviolet Rays
D022762:CCAAT-Enhancer-Binding Proteins; D002457:Cell Extracts; D004268:DNA-Binding Proteins; D007148:Immunoglobulin mu-Chains; D007476:Ionophores; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D009838:Oligodeoxyribonucleotides; D014157:Transcription Factors; C062477:endoplasmic reticulum glycoprotein p72; D000001:Calcimycin; D004247:DNA