Abstract
Chromosome segregation in mitosis is orchestrated by the interaction of the kinetochore with spindle microtubules. Our recent study shows that NEK2A interacts with MAD1 at the kinetochore and possibly functions as a novel integrator of spindle checkpoint signaling. However, it is unclear how NEK2A regulates kinetochore-microtubule attachment in mitosis. Here we show that NEK2A phosphorylates human Sgo1 and such phosphorylation is essential for faithful chromosome congression in mitosis. NEK2A binds directly to HsSgo1 in vitro and co-distributes with HsSgo1 to the kinetochore of mitotic cells. Our in vitro phosphorylation experiment demonstrated that HsSgo1 is a substrate of NEK2A and the phosphorylation sites were mapped to Ser14 and Ser507 as judged by the incorporation of 32P. Although such phosphorylation is not required for assembly of HsSgo1 to the kinetochore, expression of non-phosphorylatable mutant HsSgo1 perturbed chromosome congression and resulted in a dramatic increase in microtubule attachment errors, including syntelic and monotelic attachments. These findings reveal a key role for the NEK2A-mediated phosphorylation of HsSgo1 in orchestrating dynamic kinetochore-microtubule interaction. We propose that NEK2A-mediated phosphorylation of human Sgo1 provides a link between centromeric cohesion and spindle microtubule attachment at the kinetochores.
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References
Cleveland DW, Mao Y, Sullivan KF . Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling. Cell 2003; 112:407–421.
Nicklas RB . The motor for poleward chromosome movement in anaphase is in or near the kinetochore. J Cell Biol 1989; 109:2245–2255.
Rieder CL, Alexander SP . Kinetochores are transported poleward along a single astral microtubule during chromosome attachment to the spindle in newt lung cells. J Cell Biol 1990; 110:81–95.
Yao X, Abrieu A, Zheng Y, et al. CENP-E forms a link between attachment of spindle microtubules to kinetochores and the mitotic checkpoint. Nat Cell Biol 2000; 2:484–491.
Kitajima TS, Kawashima SA, Watanabe Y . The conserved kinetochore protein shugoshin protects centromeric cohesion during meiosis. Nature 2004; 427:510–517.
Salic A, Waters JC, Mitchison TJ . Vertebrate shugoshin links sister centromere cohesion and kinetochore microtubule stability in mitosis. Cell 2004; 118:567–578.
McGuinness BE, Hirota T, Kudo NR, et al. Shugoshin prevents dissociation of cohesin from centromeres during mitosis in vertebrate cells. PLoS Biol 2005; 3:e86.
Katis VL, Galova M, Rabitsch KP, et al. Maintenance of cohesin at centromeres after meiosis I in budding yeast requires a kinetochore-associated protein related to MEI-S332. Curr Biol 2005; 14:560–572.
Marston AL, Tham WH, Shah H, et al. A genome-wide screen identifies genes required for centromeric cohesion. Science 2004; 303:1367–1370.
Rabitsch KP, Gregan J, Schleiffer A, et al. Two fission yeast homologs of Drosophila Mei-S332 are required for chromosome segregation during meiosis I and II. Curr Biol 2004; 14:287–301.
Nigg EA . Mitotic kinases as regulators of cell division and its checkpoints. Nat Rev Mol Cell Biol 2001; 2:21–32.
O'Connell MJ, Krien MJ, Hunter T . Never say never again. The NIMA-related protein kinases in mitotic control. Trends Cell Biol 2003; 13:221–228.
Lou Y, Yao J, Zereshki A, et al. NEK2A interacts with MAD1 and possibly functions as a novel integrator of the spindle checkpoint signaling. J Biol Chem 2004; 279:20049–20057.
Zhou R, Cao X, Watson C, et al. Characterization of protein kinase A-mediated phosphorylation of ezrin in gastric parietal cell activation. J Biol Chem 2003; 278:35651–35659.
Yao X, Anderson KL, Cleveland DW . The microtubule-dependent motor centromere-associated protein E (CENP-E) is an integral component of kinetochore corona fibers that link centromeres to spindle microtubules. J Cell Biol 1997; 139:435–447.
Xue Y, Zhou F, Lu H, Chen G, Yao X . GPS: a comprehensive www server for phosphorylation sites prediction. Nucl Acids Res 2005; 33:W184–W187.
Fu G, Hua S, Ward T, et al. D-box is required for the degradation of human Shugoshin and chromosome alignment. Biochem Biophy Res Commun 2007; 357:672–678.
Tang Z, Song Y, Harley SE, et al. Human Bub1 protects centromeric sister-chromatid cohesion through Shugoshin during mitosis. Proc Natl Acad Sci USA 2004; 101:18012–18017.
Waizenegger IC, Hauf S, Meinke A, Peters JM . Two distinct pathways remove mammalian cohesin from chromosome arms in prophase and from centromeres in anaphase. Cell 2005; 103:399–410.
Church K, Lin HP . Kinetochore microtubules and chromosome movement during prometaphase in Drosphila melanogaster spermatocytes studied in life and with the electron microscope. Chromosoma 1985; 92:273–282.
Ault JG, Nicklas RB . Tension, microtubule rearrangements, and the proper distribution of chromosomes in mitosis. Chromosoma 1989; 98:33–39.
Acknowledgements
We thank members of our group for insightful discussion during the course of this study. This work was supported by grants from Chinese Academy of Science (KSCX1-YW-R65; KSCX2-YW-H10), National Basic Research Program of China (2002CB713700), Hi-Tech Research and Development Program of China (2001AA215331), Chinese Minister of Education (20020358051 to XY; PCSIRT0413 to XD), National Natural Science Foundation of China (39925018; 30270293 to XY; 30500183 to XD; 30600222 to JY), and National Institutes of Health (USA) (DK56292; CA92080) to XY (a Georgia Cancer Coalition Eminent Scholar). JY was supported by China Postdoctor (2005037560).
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Fu, G., Ding, X., Yuan, K. et al. Phosphorylation of human Sgo1 by NEK2A is essential for chromosome congression in mitosis. Cell Res 17, 608–618 (2007). https://doi.org/10.1038/cr.2007.55
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DOI: https://doi.org/10.1038/cr.2007.55
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