Tight association of DNA polymerase alpha with granular structures in the nuclear matrix of chick embryo cell: immunocytochemical detection with monoclonal antibody against DNA polymerase alpha.
Immunofluorescent methods using a monoclonal antibody against chick DNA polymerase alpha and a rabbit antibody against chick DNA polymerase beta demonstrated that both DNA polymerases alpha and beta are present mainly in nuclei of cultured chick embryo cells. Fluorescence produced by anti-DNA polymerase alpha was more intense in the small granules than in other parts of the nucleus but, fluorescence produced by anti-DNA polymerase beta was distributed evenly in the nucleus. Cells first were treated with Nonidet P-40, followed by treatment with 50 micrograms/ml pancreatic DNase and 2 M NaCl in order to prepare the nuclear matrix. Fluorescence produced by anti-DNA polymerase alpha was still detectable in the granules after these treatments, but most of the fluorescence produced by anti-DNA polymerase beta disappeared. Our results indicate that a part of DNA polymerase alpha is tightly bound to a special structure present in the nuclear matrix which presumably is the DNA replication machinery.
D000818:Animals; D000911:Antibodies, Monoclonal; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D002642:Chick Embryo; D004256:DNA Polymerase I / Q000378:metabolism*; D004257:DNA Polymerase II / Q000378:metabolism*; D003850:Deoxyribonuclease I; D003902:Detergents; D004706:Endodeoxyribonucleases / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D003902:Detergents; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D017830:Octoxynol; C010615:Nonidet P-40; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I