The B1C8 protein is in the dense assemblies of the nuclear matrix and relocates to the spindle and pericentriolar filaments at mitosis.
The B1C8 monoclonal antibody detects a 180-kDa nuclear matrix-specific protein. The protein is a component of the dense, metabolically active bodies or assemblies revealed by resinless section electron microscopy of the nuclear matrix. These assemblies are scattered through the nuclear interior, enmeshed in a complex network of 11-nm filaments. Resinless section electron microscopy of immunogold-stained nuclear matrix preparations shows B1C8 located in many but apparently not all the assemblies. In this regard, the B1C8 antigen resembles previously studied nuclear matrix proteins such as the H1B2 protein. The speckled pattern of nuclear immunofluorescence by B1C8 reflects this labeling of the dense assemblies in the nuclear matrix. Somewhat unusual is the faint staining of cytoplasmic microtubules by B1C8, which appears to be due to a weakly cross-reacting protein. During cell division, the B1C8 antigen redistributed drastically, showing the dispersion of nuclear matrix assemblies at mitosis. Speckles of B1C8 fluorescence first coalesced at prophase within the nuclear interior and then scattered into numerous cytoplasmic speckles by prometaphase. At metaphase, the B1C8 speckled cytoplasmic staining had become even more widely distributed and finely grained. Also, intense labeling appeared at the mitotic pole and on the spindle fibers themselves. The reassembly of B1C8 antigens into larger cytoplasmic speckles began at anaphase and finally, at telophase, most B1C8 labeling redistributed into speckles in the re-forming nuclei.
Wan
K M
KM
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Nickerson
J A
JA
Krockmalnic
G
G
Penman
S
S
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D002460:Cell Line; D002502:Centrioles / Q000378:metabolism / Q000648:ultrastructure; D003429:Cross Reactions; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D007399:Interphase; D016253:Microscopy, Immunoelectron; D008869:Microtubule-Associated Proteins / Q000276:immunology; D008938:Mitosis; D015530:Nuclear Matrix / Q000378:metabolism* / Q000648:ultrastructure; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D034961:Antigens, Nuclear; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins