Cytochemical evidence for the presence of actin in the nucleus of the voodoo lily appendix |
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Authors: | Skubatz H Orellana M V Yablonka-Reuveni Z |
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Institution: | (1) Department of Botany, University of Washington, Seattle, WA 98195, USA;(2) Department of Bioengineering, University of Washington, Seattle, WA 98195, USA;(3) Department of Biological Structure, University of Washington, Seattle, WA 98195, USA |
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Abstract: | Immunoflorescence microscopy of sections of the voodoo lily Sauromatum guttatum appendix stained with monoclonal antibodies against -smooth muscle actin and cytoplasmic actin revealed different staining intensity of different parts of the cell. The anti-cytoplasmic-actin recognized antigens present mainly in the cytoplasm, and the anti- -smooth muscle-actin recognized more intensively antigens present in the nuclei. A positive staining of the nucleus was also obtained with FITC–phalloidin confirming the presence of actin in its filamenous form in the nucleus. The presence of a nuclear -smooth muscle-actin-like protein was further confirmed by confocal laser microscopy. On Western blots, the two anti-actins labelled a protein band that comigrated with standard actin at the approximate molecular weight of 43 kDa. Several other proteins interacted with the two antibodies to a different degree. The monoclonal antibodies against -tubulin subunit stained only the periphery of the cytoplasm and anti-pan cytoplasmic myosin stained the cytoplasm weakly. On a Western blot, anti- -tubulin subunit primarily recognized a protein band at the appropriate molecular weight of 50 kDa. This is the first cytochemical evidence for the presence of -smooth muscle-actin-like protein in the plant nucleus. |
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