1887

Abstract

Rho GTPases are small proteins present in all eukaryotic cells, from yeast to mammals, with a function in actin organization and morphogenetic processes. Rho4 is not essential but it displays a role during cell separation at high temperature. In fact, Rho4 is involved in the secretion of the hydrolytic enzymes that are required for cell septum degradation during this process. In Δ cells, vesicles accumulate in the septum area and the glucanases Eng1 and Agn1 are not secreted to the culture medium. The localization of Eng1 and Agn1 depends on the exocyst and the septins. The exocyst is a conserved multiprotein complex important for the targeting and fusion of Golgi-derived vesicles with the plasma membrane. Septins are a family of GTP-binding proteins conserved in eukaryotes that function during cytokinesis. Here we show that Rho4 is required for the proper localization of the exocyst and septins at high temperature. Moreover, pull-down experiments demonstrate that Rho4 can interact with exocyst subunits, such as Sec8 and Exo70, and septin proteins, such as Spn3. We observe that Sec8 preferentially binds to activated GTP-Rho4, suggesting that Sec8 could be an effector of this GTPase. We propose that the interaction of Rho4 with the exocyst and septins confers a precise regulation for the secretion of glucanases at the appropriate place and time during the cell cycle.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000062
2015-05-01
2024-12-14
Loading full text...

Full text loading...

/deliver/fulltext/micro/161/5/948.html?itemId=/content/journal/micro/10.1099/mic.0.000062&mimeType=html&fmt=ahah

References

  1. An H., Morrell J. L., Jennings J. L., Link A. J., Gould K. L. 2004; Requirements of fission yeast septins for complex formation, localization, and function. Mol Biol Cell 15:5551–5564 [View Article][PubMed]
    [Google Scholar]
  2. Andrews P. D., Stark M. J. 2000; Type 1 protein phosphatase is required for maintenance of cell wall integrity, morphogenesis and cell cycle progression in Saccharomyces cerevisiae. . J Cell Sci 113:507–520[PubMed]
    [Google Scholar]
  3. Arellano M., Durán A., Pérez P. 1997; Localisation of the Schizosaccharomyces pombe rho1p GTPase and its involvement in the organisation of the actin cytoskeleton. J Cell Sci 110:2547–2555[PubMed]
    [Google Scholar]
  4. Bendezú F. O., Martin S. G. 2011; Actin cables and the exocyst form two independent morphogenesis pathways in the fission yeast. Mol Biol Cell 22:44–53 [View Article][PubMed]
    [Google Scholar]
  5. Bendezú F. O., Vincenzetti V., Martin S. G. 2012; Fission yeast Sec3 and Exo70 are transported on actin cables and localize the exocyst complex to cell poles. PLoS ONE 7:e40248 [View Article][PubMed]
    [Google Scholar]
  6. Dong Y., Pruyne D., Bretscher A. 2003; Formin-dependent actin assembly is regulated by distinct modes of Rho signaling in yeast. J Cell Biol 161:1081–1092 [View Article][PubMed]
    [Google Scholar]
  7. Dünkler A., Wendland J. 2007; Candida albicans Rho-type GTPase-encoding genes required for polarized cell growth and cell separation. Eukaryot Cell 6:844–854 [View Article][PubMed]
    [Google Scholar]
  8. Elias M., Klimes V. 2012; Rho GTPases deciphering the evolutionary history of a complex protein family. Methods Mol Biol 827:13–34
    [Google Scholar]
  9. Estravís M., Rincón S. A., Santos B., Pérez P. 2011; Cdc42 regulates multiple membrane traffic events in fission yeast. Traffic 12:1744–1758 [View Article][PubMed]
    [Google Scholar]
  10. Estravís M., Rincón S., Pérez P. 2012; Cdc42 regulation of polarized traffic in fission yeast. Commun Integr Biol 5:370–373 [View Article][PubMed]
    [Google Scholar]
  11. Forsburg S. L., Sherman D. A. 1997; General purpose tagging vectors for fission yeast. Gene 191:191–195 [View Article][PubMed]
    [Google Scholar]
  12. Free S. J. 2013; Fungal cell wall organization and biosynthesis. Adv Genet 81:33–82 [View Article][PubMed]
    [Google Scholar]
  13. García I., Jiménez D., Martín V., Durán A., Sánchez Y. 2005; The alpha-glucanase Agn1p is required for cell separation in Schizosaccharomyces pombe. . Biol Cell 97:569–576 [View Article][PubMed]
    [Google Scholar]
  14. García P., Tajadura V., García I., Sánchez Y. 2006; Role of Rho GTPases and Rho-GEFs in the regulation of cell shape and integrity in fission yeast. Yeast 23:1031–1043 [View Article][PubMed]
    [Google Scholar]
  15. Goyal A., Takaine M., Simanis V., Nakano K. 2011; Dividing the spoils of growth and the cell cycle: The fission yeast as a model for the study of cytokinesis. Cytoskeleton (Hoboken) 68:69–88 [View Article][PubMed]
    [Google Scholar]
  16. Heider M. R., Munson M. 2012; Exorcising the exocyst complex. Traffic 13:898–907 [View Article][PubMed]
    [Google Scholar]
  17. Hsu S. C., Hazuka C. D., Roth R., Foletti D. L., Heuser J., Scheller R. H. 1998; Subunit composition, protein interactions, and structures of the mammalian brain sec6/8 complex and septin filaments. Neuron 20:1111–1122 [View Article][PubMed]
    [Google Scholar]
  18. Ito H., Fukuda Y., Murata K., Kimura A. 1983; Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168[PubMed]
    [Google Scholar]
  19. Iwaki N., Karatsu K., Miyamoto M. 2003; Role of guanine nucleotide exchange factors for Rho family GTPases in the regulation of cell morphology and actin cytoskeleton in fission yeast. Biochem Biophys Res Commun 312:414–420 [View Article][PubMed]
    [Google Scholar]
  20. Jourdain I., Dooley H. C., Toda T. 2012; Fission yeast sec3 bridges the exocyst complex to the actin cytoskeleton. Traffic 13:1481–1495 [View Article][PubMed]
    [Google Scholar]
  21. Kita A., Li C., Yu Y., Umeda N., Doi A., Yasuda M., Ishiwata S., Taga A., Horiuchi Y., Sugiura R. 2011; Role of the Small GTPase Rho3 in Golgi/Endosome trafficking through functional interaction with adaptin in Fission Yeast. PLoS ONE 6:e16842 [View Article][PubMed]
    [Google Scholar]
  22. Kwon M. J., Arentshorst M., Roos E. D., van den Hondel C. A., Meyer V., Ram A. F. 2011; Functional characterization of Rho GTPases in Aspergillus niger uncovers conserved and diverged roles of Rho proteins within filamentous fungi. Mol Microbiol 79:1151–1167 [View Article][PubMed]
    [Google Scholar]
  23. Longtine M. S., Bi E. 2003; Regulation of septin organization and function in yeast. Trends Cell Biol 13:403–409 [View Article][PubMed]
    [Google Scholar]
  24. Longtine M. S., DeMarini D. J., Valencik M. L., Al-Awar O. S., Fares H., De Virgilio C., Pringle J. R. 1996; The septins: roles in cytokinesis and other processes. Curr Opin Cell Biol 8:106–119 [View Article][PubMed]
    [Google Scholar]
  25. Martín-Cuadrado A. B., Dueñas E., Sipiczki M., Vázquez de Aldana C. R., del Rey F. 2003; The endo-beta-1,3-glucanase eng1p is required for dissolution of the primary septum during cell separation in Schizosaccharomyces pombe . J Cell Sci 116:1689–1698 [View Article][PubMed]
    [Google Scholar]
  26. Martín-Cuadrado A. B., Morrell J. L., Konomi M., An H., Petit C., Osumi M., Balasubramanian M., Gould K. L., Del Rey F., de Aldana C. R. 2005; Role of septins and the exocyst complex in the function of hydrolytic enzymes responsible for fission yeast cell separation. Mol Biol Cell 16:4867–4881 [View Article][PubMed]
    [Google Scholar]
  27. Matsui Y., Toh-E A. 1992b). Yeast RHO3 and RHO4 ras superfamily genes are necessary for bud growth, and their defect is suppressed by a high dose of bud formation genes CDC42 and BEM1. . Mol Cell Biol 12:5690–5699[PubMed]
    [Google Scholar]
  28. Moreno S., Klar A., Nurse P. 1991; Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. . Methods Enzymol 194:795–823 [View Article][PubMed]
    [Google Scholar]
  29. Muñoz S., Manjón E., Sánchez Y. 2014; The putative exchange factor Gef3p interacts with Rho3p GTPase and the septin ring during cytokinesis in fission yeast. J Biol Chem 289:21995–22007 [View Article][PubMed]
    [Google Scholar]
  30. Nakano K., Mutoh T., Arai R., Mabuchi I. 2003; The small GTPase Rho4 is involved in controlling cell morphology and septation in fission yeast. Genes Cells 8:357–370 [View Article][PubMed]
    [Google Scholar]
  31. Oh Y., Bi E. 2011; Septin structure and function in yeast and beyond. Trends Cell Biol 21:141–148 [View Article][PubMed]
    [Google Scholar]
  32. Pérez P., Rincón S. A. 2010; Rho GTPases: regulation of cell polarity and growth in yeasts. Biochem J 426:243–253 [View Article][PubMed]
    [Google Scholar]
  33. Rasmussen C. G., Glass N. L. 2005; A Rho-type GTPase, rho-4, is required for septation in Neurospora crassa. . Eukaryot Cell 4:1913–1925 [View Article][PubMed]
    [Google Scholar]
  34. Robinson N. G., Guo L., Imai J., Toh-E A., Matsui Y., Tamanoi F. 1999; Rho3 of Saccharomyces cerevisiae, which regulates the actin cytoskeleton and exocytosis, is a GTPase which interacts with Myo2 and Exo70. Mol Cell Biol 19:3580–3587[PubMed]
    [Google Scholar]
  35. Roncero C., Sánchez Y. 2010; Cell separation and the maintenance of cell integrity during cytokinesis in yeast: the assembly of a septum. Yeast 27:521–530 [View Article][PubMed]
    [Google Scholar]
  36. Santos B., Gutiérrez J., Calonge T. M., Pérez P. 2003; Novel Rho GTPase involved in cytokinesis and cell wall integrity in the fission yeast Schizosaccharomyces pombe. . Eukaryot Cell 2:521–533 [View Article][PubMed]
    [Google Scholar]
  37. Santos B., Martín-Cuadrado A. B., Vázquez de Aldana C. R., del Rey F., Pérez P. 2005; Rho4 GTPase is involved in secretion of glucanases during fission yeast cytokinesis. Eukaryot Cell 4:1639–1645 [View Article][PubMed]
    [Google Scholar]
  38. Sharifmoghadam M. R., de Leon N., Hoya M., Curto M. A., Valdivieso M. H. 2010; Different steps of sexual development are differentially regulated by the Sec8p and Exo70p exocyst subunits. FEMS Microbiol Lett 305:71–80 [View Article][PubMed]
    [Google Scholar]
  39. Tasto J. J., Morrell J. L., Gould K. L. 2003; An anillin homologue, Mid2p, acts during fission yeast cytokinesis to organize the septin ring and promote cell separation. J Cell Biol 160:1093–1103 [View Article][PubMed]
    [Google Scholar]
  40. Wang H., Tang X., Liu J., Trautmann S., Balasundaram D., McCollum D. B., Balasubramanian M. K. 2002; The multiprotein exocyst complex is essential for cell separation in Schizosaccharomyces pombe. . Mol Biol Cell 13:515–529 [View Article][PubMed]
    [Google Scholar]
  41. Wang H., Tang X., Balasubramanian M. K. 2003; Rho3p regulates cell separation by modulating exocyst function in Schizosaccharomyces pombe. . Genetics 164:1323–1331[PubMed]
    [Google Scholar]
  42. Wang N., Wang M., Zhu Y. H., Grosel T. W., Sun D., Kudryashov D. S., Wu J. Q. 2015; The Rho-GEF Gef3 interacts with the septin complex and activates the GTPase Rho4 during fission yeast cytokinesis. Mol Biol Cell 26:238–255 [View Article][PubMed]
    [Google Scholar]
  43. Wu H., Turner C., Gardner J., Temple B., Brennwald P. 2010; The Exo70 subunit of the exocyst is an effector for both Cdc42 and Rho3 function in polarized exocytosis. Mol Biol Cell 21:430–442 [View Article][PubMed]
    [Google Scholar]
  44. Yu Y., Li C., Kita A., Katayama Y., Kubouchi K., Udo M., Imanaka Y., Ueda S., Masuko T., Sugiura R. 2013; Sip1, an AP-1 accessory protein in fission yeast, is required for localization of Rho3 GTPase. PLoS ONE 8:e68488 [View Article][PubMed]
    [Google Scholar]
  45. Zhu Y. H., Ye Y., Wu Z., Wu J. Q. 2013; Cooperation between Rho-GEF Gef2 and its binding partner Nod1 in the regulation of fission yeast cytokinesis. Mol Biol Cell 24:3187–3204 [View Article][PubMed]
    [Google Scholar]
/content/journal/micro/10.1099/mic.0.000062
Loading
/content/journal/micro/10.1099/mic.0.000062
Loading

Data & Media loading...

Supplements

Supplementary Data

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error