Novel nuclear and cytoplasmic proteins detected by anti-Zoothamnium arbuscula (Protozoa) spasmin 1 antibody in mammalian cells are dependent on the cell cycle.
Spasmin is a calcium-binding protein that is the major component of calcium-induced contractile filaments, called spasmoneme, found in vorticellid ciliates. Such filaments have not been observed in any organisms other than green algae. To determine whether calcium-induced contractile filaments resembling spasmoneme are present in higher eukaryotes, we performed immunofluorescence imaging of an anti-Zoothamnium arbuscula (protozoa, ciliophora) spasmin 1 polyclonal antibody in HeLa cells. In the cytoplasm, ubiquitous antigens seemed to be co-localized with microtubules at interphase, but not throughout mitosis. In the nucleus, areas linked to the nuclear envelope contained a number of hot spots. These regions were unclear during condensation of the replicated chromosomes, but became clearly visible again at cytokinesis. Immunoblotting analysis identified localized antigens during different phases of the cell cycle, including a 68/71 kDa cytoplasmic protein and a 55 kDa nuclear protein in interphase, and a 55/70 kDa protein in mitosis. The anti-spasmin 1 antibody recognized antigens in both hamster kidney BHK21 cells and Human lung cancer A-549 cells. These results suggest that novel spasmin-like proteins could be common in mammalian cells.
Itabashi
Takeshi
T
Major in Integrative Bioscience and Biomedical Engineering, Graduate School of Science and Engineering, Waseda University, Tokyo, 169-8555, Japan. t.itabashi@ruri.waseda.jp
Terasaki
Takeo
T
Asai
Hiroshi
H
D000818:Animals; D000913:Antibodies, Protozoan / Q000737:chemistry*; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000737:chemistry* / Q000276:immunology; D002118:Calcium / Q000737:chemistry; D002453:Cell Cycle / Q000502:physiology*; D002460:Cell Line; D016798:Ciliophora; D003285:Contractile Proteins / Q000276:immunology*; D006224:Cricetinae; D003593:Cytoplasm / Q000737:chemistry*; D005455:Fluorescent Antibody Technique / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000737:chemistry*; D015800:Protozoan Proteins / Q000276:immunology*
D000913:Antibodies, Protozoan; D000936:Antigen-Antibody Complex; D003285:Contractile Proteins; D009687:Nuclear Proteins; D015800:Protozoan Proteins; C026555:spasmin; D002118:Calcium