Phosphorylation of myosin II-interacting guanine nucleotide exchange factor (MyoGEF) at threonine 544 by aurora B kinase promotes the binding of polo-like kinase 1 to MyoGEF.
We previously reported that phosphorylation of myosin II-interacting guanine nucleotide exchange factor (MyoGEF) by polo-like kinase 1 (Plk1) promotes the localization of MyoGEF to the central spindle and increases MyoGEF activity toward RhoA during mitosis. In this study we report that aurora B-mediated phosphorylation of MyoGEF at Thr-544 creates a docking site for Plk1, leading to the localization and activation of MyoGEF at the central spindle. In vitro kinase assays show that aurora B can phosphorylate MyoGEF. T544A mutation drastically decreases aurora B-mediated phosphorylation of MyoGEF in vitro and in transfected HeLa cells. Coimmunoprecipitation and in vitro pulldown assays reveal that phosphorylation of MyoGEF at Thr-544 enhances the binding of Plk1 to MyoGEF. Immunofluorescence analysis shows that aurora B colocalizes with MyoGEF at the central spindle and midbody during cytokinesis. Suppression of aurora B activity by an aurora B inhibitor disrupts the localization of MyoGEF to the central spindle. In addition, T544A mutation interferes with the localization of MyoGEF to the cleavage furrow and decreases MyoGEF activity toward RhoA during mitosis. Taken together, our results suggest that aurora B coordinates with Plk1 to regulate MyoGEF activation and localization, thus contributing to the regulation of cytokinesis.
Department of Biological Sciences, Fordham University, Bronx, New York 10458.
Di
D
Wu
Department of Surgery, College of Medicine, Mayo Clinic Rochester, Minnesota 55902, and.
Michael
M
Asiedu
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey 08855.
Fumio
F
Matsumura
Department of Biological Sciences, Fordham University, Bronx, New York 10458. Electronic address: qwei3@fordham.edu.
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Wei
D064107:Aurora Kinase B / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D048749:Cytokinesis; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D008938:Mitosis; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism; D013912:Threonine / Q000235:genetics / Q000378:metabolism*; D000097869:Polo-Like Kinase 1
D064107:Aurora Kinase B; D018797:Cell Cycle Proteins; D020662:Guanine Nucleotide Exchange Factors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D013912:Threonine; D000097869:Polo-Like Kinase 1; C527325:PLEKHG6 protein, human; C576921:AURKB protein, human
Aurora B Kinase; Central Spindle; Cytokinesis; Guanine Nucleotide Exchange Factor (GEF); Mitosis; MyoGEF; Phosphorylation; Plk1; Rho GTPases