The Rsr1/Bud1 GTPase Interacts with Itself and the Cdc42 GTPase During Bud-site Selection and Polarity Establishment in Budding Yeast

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2010 Jul 1

PMID 20587777

Citations 31

Authors

Pil Jung Kang

Laure Beven

Seethalakshmi Hariharan

Hay-Oak Park

Affiliations

Soon will be listed here.

Abstract

Cell polarization occurs along a single axis that is generally determined in response to spatial cues. In budding yeast, the Rsr1 GTPase and its regulators direct the establishment of cell polarity at the proper cortical location in response to cell type-specific cues. Here we use a combination of in vivo and in vitro approaches to understand how Rsr1 polarization is established. We find that Rsr1 associates with itself in a spatially and temporally controlled manner. The homotypic interaction and localization of Rsr1 to the mother-bud neck and to the subsequent division site are dependent on its GDP-GTP exchange factor Bud5. Analyses of rsr1 mutants suggest that Bud5 recruits Rsr1 to these sites and promotes the homodimer formation. Rsr1 also exhibits heterotypic interaction with the Cdc42 GTPase in vivo. We show that the polybasic region of Rsr1 is necessary for the efficient homotypic and heterotypic interactions, selection of a proper growth site, and polarity establishment. Our findings thus suggest that dimerization of GTPases may be an efficient mechanism to set up cellular asymmetry.

Citing Articles

Gradient tracking in mating yeast depends on Bud1 inactivation and actin-independent vesicle delivery.

Wang X, Pai C, Stone D J Cell Biol. 2022; 221(12).

PMID: 36156058 PMC: 9516845. DOI: 10.1083/jcb.202203004.

The Path towards Predicting Evolution as Illustrated in Yeast Cell Polarity.

Daalman W, Sweep E, Laan L Cells. 2020; 9(12).

PMID: 33255231 PMC: 7760196. DOI: 10.3390/cells9122534.

Overexpression of the Small GTPase, RsrA, Promotes Polarity Establishment during Germination.

Martin-Vicente A, Souza A, Nywening A, Ge W, Fortwendel J J Fungi (Basel). 2020; 6(4).

PMID: 33202962 PMC: 7711769. DOI: 10.3390/jof6040285.

Activation of Cdc42 GTPase upon CRY2-Induced Cortical Recruitment Is Antagonized by GAPs in Fission Yeast.

Lamas I, Weber N, Martin S Cells. 2020; 9(9).

PMID: 32932721 PMC: 7565336. DOI: 10.3390/cells9092089.

Regulation of Cdc42 for polarized growth in budding yeast.

Miller K, Kang P, Park H Microb Cell. 2020; 7(7):175-189.

PMID: 32656257 PMC: 7328677. DOI: 10.15698/mic2020.07.722.

References

Etienne-Manneville S . Cdc42--the centre of polarity. J Cell Sci. 2004; 117(Pt 8):1291-300. DOI: 10.1242/jcs.01115. View

Pringle J . Staining of bud scars and other cell wall chitin with calcofluor. Methods Enzymol. 1991; 194:732-5. DOI: 10.1016/0076-6879(91)94055-h. View

Kozminski K, Beven L, Angerman E, Tong A, Boone C, Park H . Interaction between a Ras and a Rho GTPase couples selection of a growth site to the development of cell polarity in yeast. Mol Biol Cell. 2003; 14(12):4958-70. PMC: 284798. DOI: 10.1091/mbc.e03-06-0426. View

Zheng Y, Bender A, Cerione R . Interactions among proteins involved in bud-site selection and bud-site assembly in Saccharomyces cerevisiae. J Biol Chem. 1995; 270(2):626-30. DOI: 10.1074/jbc.270.2.626. View

Zhang B, Gao Y, Moon S, Zhang Y, Zheng Y . Oligomerization of Rac1 gtpase mediated by the carboxyl-terminal polybasic domain. J Biol Chem. 2001; 276(12):8958-67. DOI: 10.1074/jbc.M008720200. View

Sung M, Huh W . Bimolecular fluorescence complementation analysis system for in vivo detection of protein-protein interaction in Saccharomyces cerevisiae. Yeast. 2007; 24(9):767-75. DOI: 10.1002/yea.1504. View

Beck R, Sun Z, Adolf F, Rutz C, Bassler J, Wild K . Membrane curvature induced by Arf1-GTP is essential for vesicle formation. Proc Natl Acad Sci U S A. 2008; 105(33):11731-6. PMC: 2575275. DOI: 10.1073/pnas.0805182105. View

Park H, Chant J, Herskowitz I . BUD2 encodes a GTPase-activating protein for Bud1/Rsr1 necessary for proper bud-site selection in yeast. Nature. 1993; 365(6443):269-74. DOI: 10.1038/365269a0. View

Zhang B, Zheng Y . Negative regulation of Rho family GTPases Cdc42 and Rac2 by homodimer formation. J Biol Chem. 1998; 273(40):25728-33. DOI: 10.1074/jbc.273.40.25728. View

10.

Cole K, McLaughlin H, Johnson D . Use of bimolecular fluorescence complementation to study in vivo interactions between Cdc42p and Rdi1p of Saccharomyces cerevisiae. Eukaryot Cell. 2007; 6(3):378-87. PMC: 1828923. DOI: 10.1128/EC.00368-06. View

11.

Krappmann A, Taheri N, Heinrich M, Mosch H . Distinct domains of yeast cortical tag proteins Bud8p and Bud9p confer polar localization and functionality. Mol Biol Cell. 2007; 18(9):3323-39. PMC: 1951750. DOI: 10.1091/mbc.e06-10-0899. View

12.

Kerppola T . Bimolecular fluorescence complementation: visualization of molecular interactions in living cells. Methods Cell Biol. 2007; 85:431-70. PMC: 2829325. DOI: 10.1016/S0091-679X(08)85019-4. View

13.

Chant J, Corrado K, Pringle J, Herskowitz I . Yeast BUD5, encoding a putative GDP-GTP exchange factor, is necessary for bud site selection and interacts with bud formation gene BEM1. Cell. 1991; 65(7):1213-24. DOI: 10.1016/0092-8674(91)90016-r. View

14.

Brown J, Jaquenoud M, Gulli M, Chant J, Peter M . Novel Cdc42-binding proteins Gic1 and Gic2 control cell polarity in yeast. Genes Dev. 1997; 11(22):2972-82. PMC: 316694. DOI: 10.1101/gad.11.22.2972. View

15.

Inouye K, Mizutani S, Koide H, Kaziro Y . Formation of the Ras dimer is essential for Raf-1 activation. J Biol Chem. 2000; 275(6):3737-40. DOI: 10.1074/jbc.275.6.3737. View

16.

Park H, Bi E . Central roles of small GTPases in the development of cell polarity in yeast and beyond. Microbiol Mol Biol Rev. 2007; 71(1):48-96. PMC: 1847380. DOI: 10.1128/MMBR.00028-06. View

17.

Yoshida S, Bartolini S, Pellman D . Mechanisms for concentrating Rho1 during cytokinesis. Genes Dev. 2009; 23(7):810-23. PMC: 2666341. DOI: 10.1101/gad.1785209. View

18.

Bender A, Pringle J . Multicopy suppression of the cdc24 budding defect in yeast by CDC42 and three newly identified genes including the ras-related gene RSR1. Proc Natl Acad Sci U S A. 1989; 86(24):9976-80. PMC: 298625. DOI: 10.1073/pnas.86.24.9976. View

19.

Marston A, Chen T, Yang M, Belhumeur P, Chant J . A localized GTPase exchange factor, Bud5, determines the orientation of division axes in yeast. Curr Biol. 2001; 11(10):803-7. DOI: 10.1016/s0960-9822(01)00230-5. View

20.

Bailly E, Cabantous S, Sondaz D, Bernadac A, Simon M . Differential cellular localization among mitotic cyclins from Saccharomyces cerevisiae: a new role for the axial budding protein Bud3 in targeting Clb2 to the mother-bud neck. J Cell Sci. 2003; 116(Pt 20):4119-30. DOI: 10.1242/jcs.00706. View