Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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CBP7 Interferes with the Multicellular Development of Dictyostelium Cells by Inhibiting Chemoattractant-Mediated Cell Aggregation

  • Park, Byeonggyu (Department of Biology & BK21- Plus Research Team for Bioactive Control Technology, College of Natural Sciences, Chosun University) ;
  • Shin, Dong-Yeop (Department of Biology & BK21- Plus Research Team for Bioactive Control Technology, College of Natural Sciences, Chosun University) ;
  • Jeon, Taeck Joong (Department of Biology & BK21- Plus Research Team for Bioactive Control Technology, College of Natural Sciences, Chosun University)
  • Received : 2017.08.15
  • Accepted : 2017.11.06
  • Published : 2018.02.28
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Abstract

Calcium ions are involved in the regulation of diverse cellular processes. Fourteen genes encoding calcium binding proteins have been identified in Dictyostelium. CBP7, one of the 14 CBPs, is composed of 169 amino acids and contains four EF-hand motifs. Here, we investigated the roles of CBP7 in the development and cell migration of Dictyostelium cells and found that high levels of CBP7 exerted a negative effect on cells aggregation during development, possibly by inhibiting chemoattractant-directed cell migration. While cells lacking CBP7 exhibited normal development and chemotaxis similar that of wild-type cells, CBP7 overexpressing cells completely lost their chemotactic abilities to move toward increasing cAMP concentrations. This resulted in inhibition of cellular aggregation, a process required for forming multicellular organisms during development. Low levels of cytosolic free calcium were observed in CBP7 overexpressing cells, which was likely the underlying cause of their lack of chemotaxis. Our results demonstrate that CBP7 plays an important role in cell spreading and cell-substrate adhesion. cbp7 null cells showed decreased cell size and cell-substrate adhesion. The present study contributes to further understanding the role of calcium signaling in regulation of cell migration and development.

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References

  1. Andre, B., Noegel, A.A., and Schleicher, M. (1996). Dictyostelium discoideum contains a family of calmodulin-related EF-hand proteins that are developmentally regulated. FEBS Lett. 382, 198-202. https://doi.org/10.1016/0014-5793(96)00176-7
  2. Catalano, A., and O'Day, D.H. (2013). Rad53 homologue forkheadassociated kinase A (FhkA). and $Ca^{2+}$-binding protein 4a (CBP4a) are nucleolar proteins that differentially redistribute during mitosis in Dictyostelium. Cell Div. 8, 4. https://doi.org/10.1186/1747-1028-8-4
  3. Chin, D., and Means, A.R. (2000). Calmodulin: a prototypical calcium sensor. Trends Cell Biol. 10, 322-328. https://doi.org/10.1016/S0962-8924(00)01800-6
  4. Chisholm, R.L., and Firtel, R.A. (2004). Insights into morphogenesis from a simple developmental system. Nat. Rev. Mol. Cell Biol. 5, 531-541. https://doi.org/10.1038/nrm1427
  5. Clapham, D.E. (2007). Calcium signaling. Cell 131, 1047-1058. https://doi.org/10.1016/j.cell.2007.11.028
  6. Dharamsi, A., Tessarolo, D., Coukell, B., and Pun, J. (2000). CBP1 associates with the Dictyostelium cytoskeleton and is important for normal cell aggregation under certain developmental conditions. Exp. Cell Res. 258, 298-309. https://doi.org/10.1006/excr.2000.4950
  7. Dorywalska, M., Coukell, B., and Dharamsi, A. (2000). Characterization and hetereologous expression of cDNAs encoding two novel closely related Ca(2+)-binding proteins in Dictyostelium discoideum. Biochim. Biophys. Acta 1496, 356-361. https://doi.org/10.1016/S0167-4889(00)00024-0
  8. Evans, J.H., and Falke, J.J. (2007). $Ca^{2+}$ influx is an essential component of the positive-feedback loop that maintains leadingedge structure and activity in macrophages. Proc. Natl. Acad. Sci. USA 104, 16176-16181. https://doi.org/10.1073/pnas.0707719104
  9. Gifford, J.L., Walsh, M.P., and Vogel, H.J. (2007). Structures and metal-ion-binding properties of the $Ca^{2+}$-binding helix-loop-helix EFhand motifs. Biochem. J. 405, 199-221. https://doi.org/10.1042/BJ20070255
  10. Jeon, T.J., Lee, D.J., Lee, S., Weeks, G., and Firtel, R.A. (2007a). Regulation of Rap1 activity by RapGAP1 controls cell adhesion at the front of chemotaxing cells. J. Cell Biol. 179, 833-843. https://doi.org/10.1083/jcb.200705068
  11. Jeon, T.J., Lee, D.J., Merlot, S., Weeks, G., and Firtel, R.A. (2007b). Rap1 controls cell adhesion and cell motility through the regulation of myosin II. J. Cell Biol. 176, 1021-1033. https://doi.org/10.1083/jcb.200607072
  12. Jeon, T.J., Lee, S., Weeks, G., and Firtel, R.A. (2009). Regulation of Dictyostelium morphogenesis by RapGAP3. Dev. Biol. 328, 210-220. https://doi.org/10.1016/j.ydbio.2009.01.016
  13. Lee, M.R., and Jeon, T.J. (2012). Cell migration: regulation of cytoskeleton by Rap1 in Dictyostelium discoideum. J. Microbiol. 50, 555-561. https://doi.org/10.1007/s12275-012-2246-7
  14. Lee, C.H., Jeong, S.Y., Kim, B.J., Choi, C.H., Kim, J.S., Koo, B.M., Seok, Y.J., Yim, H.S., and Kang, S.O. (2005). Dictyostelium CBP3 associates with actin cytoskeleton and is related to slug migration. Biochim. Biophys. Acta 1743, 281-290. https://doi.org/10.1016/j.bbamcr.2005.01.003
  15. Lusche, D.F., Wessels, D., and Soll, D.R. (2009). The effects of extracellular calcium on motility, pseudopod and uropod formation, chemotaxis, and the cortical localization of myosin II in Dictyostelium discoideum. Cell Motil Cytoskeleton. 66, 567-587. https://doi.org/10.1002/cm.20367
  16. Malchow, D., Mutzel, R., and Schlatterer, C. (1996). On the role of calcium during chemotactic signalling and differentiation of the cellular slime mould Dictyostelium discoideum. Int. J. Dev. Biol. 40, 135-139.
  17. Mishig-Ochiriin, T., Lee, C.H., Jeong, S.Y., Kim, B.J., Choi, C.H., Yim, H.S., and Kang, S.O. (2005). Calcium-induced conformational changes of the recombinant CBP3 protein from Dictyostelium discoideum. Biochim. Biophys. Acta 1748, 157-164. https://doi.org/10.1016/j.bbapap.2004.12.018
  18. Mun, H., Lee, M.R., and Jeon, T.J. (2014). RapGAP9 regulation of the morphogenesis and development in Dictyostelium. Biochem. Biophys. Res. Commun. 446, 428-433. https://doi.org/10.1016/j.bbrc.2014.01.196
  19. Myre, M.A., and O'Day, D.H. (2004). Dictyostelium calcium-binding protein 4a interacts with nucleomorphin, a BRCT-domain protein that regulates nuclear number. Biochem. Biophys. Res. Commun. 322, 665-671. https://doi.org/10.1016/j.bbrc.2004.07.168
  20. Rot, G., Parikh, A., Curk, T., Kuspa, A., Shaulsky, G., and Zupan, B. (2009). dictyExpress: a Dictyostelium discoideum gene expression database with an explorative data analysis web-based interface. BMC Bioinformatics 10, 265. https://doi.org/10.1186/1471-2105-10-265
  21. Sakamoto, H., Nishio, K., Tomisako, M., Kuwayama, H., Tanaka, Y., Suetake, I., Tajima, S., Ogihara, S., Coukell, B., and Maeda, M. (2003). Identification and characterization of novel calcium-binding proteins of Dictyostelium and their spatial expression patterns during development. Dev. Growth Differ. 45, 507-514. https://doi.org/10.1111/j.1440-169X.2003.00718.x
  22. Siu, C.H., Sriskanthadevan, S., Wang, J., Hou, L., Chen, G., Xu, X., Thomson, A., and Yang, C. (2011). Regulation of spatiotemporal expression of cell-cell adhesion molecules during development of Dictyostelium discoideum. Dev. Growth Differ. 53, 518-527. https://doi.org/10.1111/j.1440-169X.2011.01267.x
  23. Tanaka, Y., Itakura, R., Amagai, A., and Maeda, Y. (1998). The signals for starvation response are transduced through elevated [$Ca^{2+}$]i in Dictyostelium cells. Exp Cell Res 240, 340-348. https://doi.org/10.1006/excr.1998.3947
  24. Yumura, S., Furuya, K., and Takeuchi, I. (1996). Intracellular free calcium responses during chemotaxis of Dictyostelium cells. J. Cell Sci. 109 (Pt 11), 2673-2678.