Redistribution of nuclear envelope associated antigen during the mitotic cycle.
Murine hybridomas were generated to DNA/tight binding proteins complex isolated from the residual nuclear structure following a procedure analogous to that yielding "empty" shells of nuclear envelope. A monoclonal antibody designated 2A8 was selected because of its differential immunostaining of mitotic cells of a synchronized mouse fibroblast cell culture L-929. The target antigen was rendered insoluble by a sequence of extractions of isolated nuclei of diverse cell types with detergents, urea, DNase I and alkali thus reproducing some solubility properties of proteins constituting an operationally defined residual nuclear matrix. The cognate polypeptide was localized on a subset of proteins of M(r) 58-65 kDa, 70 kDa in isolated fibroblast nuclear matrices. The functional implication of the antigen in mitosis-related disassembly-assembly process of the nuclear matrix/envelope was detected. At prophase the antibody decorated the nuclear periphery and nuclear envelope fixed inward filaments. A fibrous network of cytoplasmic localization was stained in metaphase. At anaphase the antigen was dispositioned into peripheral fibrogranular clusters of polar orientation predominantly on one side of the nucleus. Proceeding to telophase a spreading fluorescence was manifested over the entire contour of the nuclear periphery to delineate the reforming nucleus. By immunogold electron microscopy of interphase cells the antigen was identified as evenly distributed in chromatin and interchromatin regions. At initiation of chromosome condensation in mitosis the label was detected predominantly in the chromosomal area.
Batova
I N
IN
Department of Immunobiology, Institute of Biology and Immunology, Sofia, Bulgaria.
Kyurkchiev
S D
SD
Russev
R
R
Kehayov
I R
IR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000032:analysis; D015153:Blotting, Western; D005347:Fibroblasts / Q000737:chemistry / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology; D007150:Immunohistochemistry; D007399:Interphase / Q000502:physiology; D051379:Mice; D008938:Mitosis / Q000502:physiology*; D009685:Nuclear Envelope / Q000737:chemistry* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens