Nuclear localization of C-terminal domains of the kinesin-like protein MKLP-1.
The successful execution of mitosis in mammalian cells requires the activities of numerous kinesin-like proteins. The Mitotic Kinesin-Like Protein-1 (MKLP-1) localizes to the spindle equator and is believed to participate in the separation of spindle poles during anaphase B. Injection of antibodies against MKLP-1 into dividing cells results in cell cycle arrest, suggesting that MKLP-1 is essential for mitosis. To further characterize MKLP-1, constructs encoding C-terminal domains of MKLP-1 were expressed as fusions to the green fluorescent protein and localized in HeLa cells. All constructs localized to the nucleus indicating the presence of at least one nuclear localization sequence in the C-terminus of the protein. C-terminal domains of MKLP-1 expressed in insect cells also localized to the nucleus as shown by subcellular fractionation. These proteins remained tightly associated with the nucleus following both detergent and salt extraction, suggesting a tight interaction with a component of the nucleus.
Deavours
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Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, 24061, USA.
Walker
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D000818:Animals; D001692:Biological Transport; D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D003902:Detergents; D006367:HeLa Cells; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019913:Nuclear Localization Signals; D015530:Nuclear Matrix / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012492:Salts; D017384:Sequence Deletion; D018411:Spodoptera / Q000166:cytology / Q000235:genetics; D014162:Transfection
D003902:Detergents; C092935:KIF23 protein, human; D008869:Microtubule-Associated Proteins; D019913:Nuclear Localization Signals; D011993:Recombinant Fusion Proteins; D012492:Salts