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Roles of the PH, coiled-coil and SAM domains of the yeast polarity protein Boi2 in polarity-site localization and function in polarized growth

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Abstract

Boi1 and Boi2 are paralogous proteins essential for bud formation in budding yeast. So far, the domains that target Boi1/Boi2 to the polarity sites and function in bud formation are not well understood. Here, we report that a coiled-coil domain of Boi2 cooperates with the adjacent PH domain to confer Boi2’s bud-cortex localization and major function in cell growth. The PH domain portion of the PH-CC bi-domain interacts with the Rho GTPases Cdc42 and Rho3 and both interactions are independent of the GTP/GDP-bound state of each GTPase. Interestingly, high-copy RHO3 and BOI2 but not CDC42 suppressed the growth defect of RGA1-C538 overexpression and the sec15-1 mutant and this BOI2 function depends on RHO3, suggesting that Boi2 may function in the Rho3 pathway. The SAM domain of Boi2 plays an essential role in high-copy suppression of the two mutants as well as in the early bud-neck localization of Boi2. The SAM domain and the CC domain also interact homotypically. They are likely involved in the formation of Boi2-containing protein complex. Our results provide new insights in the localization and function of Boi2 and highlight the importance of the PH-CC bi-domain and the SAM domain in Boi2’s localization and function.

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Acknowledgements

We thank Drs. Erfei Bi, Nils Johnsson and Patrick Brennwald for kindly providing yeast strains and plasmids. This work was supported by the National Natural Science Foundation of China (Grant numbers 31570076 and 31870062).

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  1. Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan, 430072, China

    Zhi-Wen Jia, Si-Xu Lv, Jing Zhu, Chen-Yang Xia & Xiang-Dong Gao

  2. Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Wuhan, China

    Xiang-Dong Gao

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  1. Zhi-Wen Jia
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Correspondence to Xiang-Dong Gao.

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Fig. S1.

Expression of GFP-Boi2 segments in yeast cells. Table S1. Yeast strains used in this study. Table S2. Plasmids used in this study. Table S3. Oligonucleotides used in this study.

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Jia, ZW., Lv, SX., Zhu, J. et al. Roles of the PH, coiled-coil and SAM domains of the yeast polarity protein Boi2 in polarity-site localization and function in polarized growth. Curr Genet 66, 1101–1115 (2020). https://doi.org/10.1007/s00294-020-01093-9

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