Production of monoclonal antibodies against RNA polymerase I from nonimmunized autoimmune MRL/lpr mice and their use in rDNA transcription analysis.
A hybridoma secreting monoclonal antibodies against RNA polymerase I was produced by the fusion of myeloma cells with spleen cells from a nonimmunized MRL/lpr mouse which is known to produce autoantibodies to RNA polymerase I. The antibodies (McAb-2D11) belong to the IgG2b subclass, reacted specifically with the second largest (120 kDa) subunit of RNA polymerase I, and inhibited accurate transcription of cloned rat rDNA in a fractionated cell extract following immunoprecipitation of RNA polymerase I. McAb-2D11 did not inhibit RNA polymerase II-mediated transcription of the mouse metallothionein-I gene. Immunocytochemical procedures with biotinylated second antibody demonstrated specific immunolocalization of RNA polymerase I in the nucleus. These studies have (a) presented direct evidence that autoantibodies to functional RNA polymerase I are produced in a murine model of systemic lupus erythematosus, (b) demonstrated specificity of the monoclonal antibody for RNA polymerase I, and (c) provided a useful tool for the purification of RNA polymerase I and/or transcription factor(s) associated with RNA polymerase I.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001327:Autoimmune Diseases / Q000276:immunology*; D004275:DNA, Ribosomal / Q000032:analysis*; D007074:Immunoglobulin G / Q000276:immunology; D008114:Liver Neoplasms, Experimental / Q000201:enzymology; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D051379:Mice; D008815:Mice, Inbred Strains / Q000276:immunology*; D008817:Mice, Mutant Strains / Q000276:immunology*; D009363:Neoplasm Proteins / Q000276:immunology; D012318:RNA Polymerase I / Q000037:antagonists & inhibitors / Q000276:immunology*; D012335:RNA, Ribosomal / Q000096:biosynthesis; D014158:Transcription, Genetic
D000911:Antibodies, Monoclonal; D004275:DNA, Ribosomal; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; D012335:RNA, Ribosomal; D012318:RNA Polymerase I