Mitotic arrest in Ptk(2) cells induced by microinjection of a rabbit antiserum and affinity-purified antibodies against a 66-kDa PtK(2) cell polypeptide.
Cell division was arrested by injection of a preimmune rabbit serum, B-61, into PtK(2) cells during interphase and prometaphase. Identical results were obtained by injection of whole B-61 antiserum and of antibodies affinity-purified from the serum against a 66-kDa PtK(2) cell polypeptide. When injected into interphase cells, the antibodies arrested further development and cell division. When injected into prometaphase and metaphase cells, spindles shortened and poles moved together at a rate of 0.2-0.4 mu m/min, approximately half the rate of anaphase A chromosome movements in normally dividing PtK(2) cells. Chromosomes decondensed and cells did not reenter division. Both whole antisera and affinity-purified antibodies stained antigens diffusely localized throughout the cytoplasm in dividing and interphase cells. These results suggest that the 66-kDa antigen is a nonspindle protein that may regulate mitotic progression in PtK(2) cells.
Byrd
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Department of Biological Sciences, Mississippi State University, Starkville, USA.
Wise
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Dentler
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000941:Antigens / Q000032:analysis; D015153:Blotting, Western; D002460:Cell Line; D003593:Cytoplasm / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera; D008845:Microinjections; D008856:Microscopy, Fluorescence; D008938:Mitosis / Q000276:immunology*; D008970:Molecular Weight; D010455:Peptides / Q000276:immunology*; D011817:Rabbits; D008941:Spindle Apparatus / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000941:Antigens; D007106:Immune Sera; D010455:Peptides