Identification of a novel microtubule-associated protein that regulates microtubule organization and cytokinesis by using a GFP-screening strategy.
Microtubules play critical roles in a variety of cell processes, including mitosis, organelle transport, adhesion and migration, and the maintenance of cell polarity. Microtubule-associated proteins (MAPs) regulate the dynamic organization and stability of microtubules, often through either cell-specific or cell division stage-specific interactions. To identify novel cytoskeletal-associated proteins and peptides that regulate microtubules and other cytoskeletal and adhesive structures, we have developed a GFP cDNA screening strategy based on identifying gene products that localize to these structures. Using this approach, we have identified a novel MAP, GLFND, that shows homology to the Opitz syndrome gene product [6], localizes to a subpopulation of microtubules that are acetylated, and protects microtubules from depolymerization with nocodazole. Expression of an N-terminal deletion binds microtubules but alters their organization. During the cell cycle, GLFND dissociates from microtubules at the beginning of mitosis and then reassociates at cytokinesis. Furthermore, ectopic expression of GLFND inhibits cell division and cytokinesis in CHO cells. These observations make GLFND unique among MAPs characterized thus far.
Manabe
Ri ichiroh
Ri
Department of Cell Biology, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
Whitmore
Leanna
L
Weiss
Jonathan M
JM
Horwitz
Alan Rick
AR
D000818:Animals; D016466:CHO Cells; D002455:Cell Division / Q000502:physiology*; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D008870:Microtubules / Q000502:physiology / Q000648:ultrastructure*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D014162:Transfection; D014404:Tubulin / Q000378:metabolism
D018076:DNA, Complementary; C429034:FSD1 protein, human; D008164:Luminescent Proteins; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D014404:Tubulin; D049452:Green Fluorescent Proteins