Affinity purification and characterization of CIF, an insect immunoresponsive factor with NF-kappa B-like properties.
1. A rapid DNA affinity purification procedure was worked out for the purification of the Cecropia Immunoresponsive Factor (CIF) from the pupae of Hyalophora cecropia. 2. CIF consists of a single polypeptide chain of 65 kDa and is present as a homodimer under native conditions. 3. CIF binds to the kappa B-like sequences upstream of the H. cecropia immune genes with the following order of affinity: attacin kappa B greater than lysozyme kappa B greater than cecropin A kappa B greater than cecropin B kappa B. 4. The purified CIF also strongly binds to the kappa B sequences from both the immunoglobulin kappa light chain gene and the MHC class I gene. 5. The DNA binding of CIF can be inhibited by antisera directed against NF-kappa B-related proteins. 6. The cytoplasmic factor Cl, co-purified from the affinity column, contains two polypeptide chains, one of which has the same molecular weight as CIF.
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Department of Microbiology, University of Stockholm, Sweden.
Faye
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D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism; D017344:Genes, Insect; D015395:Histocompatibility Antigens Class I / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000494:pharmacology; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009036:Moths / Q000235:genetics / Q000276:immunology; D016328:NF-kappa B / Q000276:immunology / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011679:Pupa / Q000737:chemistry
D015395:Histocompatibility Antigens Class I; D007106:Immune Sera; D007145:Immunoglobulin kappa-Chains; D046911:Macromolecular Substances; D016328:NF-kappa B; D009687:Nuclear Proteins; D004247:DNA