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Roles of Dhh1 RNA helicase in yeast filamentous growth: Analysis of N-terminal phosphorylation residues and ATPase domains

  • Microbial Genetics, Genomics and Molecular Biology
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Abstract

In yeast Saccharomyces cerevisiae, the Dhh1 protein, a member of the DEAD-box RNA helicase, stimulates Dcp2/Dcp1-mediated mRNA decapping and functions as a general translation repressor. Dhh1 also positively regulates translation of a selected set of mRNAs, including Ste12, a transcription factor for yeast mating and pseudohyphal growth. Given the diverse functions of Dhh1, we investigated whether the putative phosphorylation sites or the conserved motifs for the DEAD-box RNA helicases were crucial in the regulatory roles of Dhh1 during pseudohyphal growth. Mutations in the ATPase A or B motif (DHH1-K96R or DHH1-D195A) showed significant defects in pseudohyphal colony morphology and agar invasive phenotypes. The N-terminal phospho-mimetic mutation, DHH1-T16E, showed defects in pseudohyphal phenotypes. Decreased levels of Ste12 protein were also observed in these pseudohyphal-defective mutant cells under filamentous-inducing low nitrogen conditions. We suggest that the ATPase motifs and the Thr16 phosphorylation site of Dhh1 are crucial to its regulatory roles in pseudohyphal growth under low nitrogen conditions.

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References

  • Albuquerque, C.P., Smolka, M.B., Payne, S.H., Bafna, V., Eng, J., and Zhou, H. 2008. A multidimensional chromatography technology for in-depth phosphoproteome analysis. Mol. Cell. Proteomics 7, 1389–1396.

    Article  CAS  Google Scholar 

  • Braus, G.H., Grundmann, O., Brückner, S., and Mösch, H.U. 2003. Amino acid starvation and Gcn4p regulate adhesive growth and FLO11 gene expression in Saccharomyces cerevisiae. Mol. Biol. Cell 14, 4272–4284.

    Article  CAS  Google Scholar 

  • Cheng, Z., Coller, J., Parker, R., and Song, H. 2005. Crystal structure and functional analysis of DEAD-box protein Dhh1p. RNA 11, 1258–1270.

    Article  CAS  Google Scholar 

  • Coller, J. and Parker, R. 2005. General translational repression by activators of mRNA decapping. Cell 122, 875–886.

    Article  CAS  Google Scholar 

  • Cullen, P.J. and Sprague, G.F. Jr. 2000. Glucose depletion causes haploid invasive growth in yeast. Proc. Natl. Acad. Sci. USA 97, 13619–13624.

    Article  CAS  Google Scholar 

  • Dutta, A., Zheng, S., Jain, D., Cameron, C.E., and Reese, J.C. 2011. Intermolecular interactions within the abundant DEAD-box protein Dhh1 regulate its activity in vivo. J. Biol. Chem. 286, 27454–27470.

    Article  CAS  Google Scholar 

  • Gancedo, J.M. 2001. Control of pseudohyphae formation in Saccharomyces cerevisiae. FEMS Microbiol. Rev. 25, 107–123.

    Article  CAS  Google Scholar 

  • Gimeno, C.J., Ljungdahl, P.O., Styles, C.A., and Fink, G.R. 1992. Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: regulation by starvation and RAS. Cell 68, 1077–1090.

    Article  CAS  Google Scholar 

  • Gnad, F., de Godoy, L.M., Cox, J., Neuhauser, N., Ren, S., Olsen, J.V., and Mann, M. 2009. High-accuracy identification and bioinformatic analysis of in vivo protein phosphorylation sites in yeast. Proteomics 9, 4642–4652.

    Article  CAS  Google Scholar 

  • He, F., Celik, A., Wu, C., and Jacobson, A. 2018. General decapping activators target different subsets of inefficiently translated mRNAs. eLife 7, e34409.

    Article  Google Scholar 

  • Jung, D., Ahn, J., Rhee, B., and Kim, J. 2017. Mutational analysis of the RNA helicase Dhh1 in Ste12 expression and yeast mating. J. Microbiol. 55, 373–378.

    Article  CAS  Google Scholar 

  • Jung, D., Seo, J.S., Nam, J., and Kim, J. 2019. Functional association of Loc1 and Puf6 with RNA helicase Dhh1 in translational regulation of Saccharomyces cerevisiae Ste12. PLoS One 14, e0220137.

    Article  CAS  Google Scholar 

  • Jungfleisch, J., Nedialkova, D.D., Dotu, I., Sloan, K.E., Martinez-Bosch, N., Brüning, L., Raineri, E., Navarro, P., Bohnsack, M.T., Leidel, S.A., et al. 2017. A novel translational control mechanism involving RNA structures within coding sequences. Genome Res. 27, 95–106.

    Article  CAS  Google Scholar 

  • Ka, M., Park, Y.U., and Kim, J. 2008. The DEAD-box RNA helicase, Dhh1, functions in mating by regulating Ste12 translation in Saccharomyces cerevisiae. Biochem. Biophys. Res. Commun. 367, 680–686.

    Article  CAS  Google Scholar 

  • Liu, H., Styles, C.A., and Fink, G.R. 1993. Elements of the yeast pheromone response pathway required for filamentous growth of diploids. Science 262, 1741–1744.

    Article  CAS  Google Scholar 

  • Liu, H., Styles, C.A., and Fink, G.R. 1996. Saccharomyces cerevisiae S288C has a mutation in FLO8, a gene required for filamentous growth. Genetics 144, 967–978.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liu, X., Yao, Z., Jin, M., Namkoong, S., Yin, Z., Lee, J.H., and Klionsky, D.J. 2019. Dhh1 promotes autophagy-related protein translation during nitrogen starvation. PLoS Biol. 17, e3000219.

    Article  CAS  Google Scholar 

  • Mugler, C.F., Hondele, M., Heinrich, S., Sachdev, R., Vallotton, P., Koek, A.Y., Chan, L.Y., and Weis, K. 2016. ATPase activity of the DEAD-box protein Dhh1 controls processing body formation. eLife 5, e18746.

    Article  Google Scholar 

  • Mutlu, N., Sheidy, D.T., Hsu, A., Jeong, H.S., Wozniak, K.J., and Kumar, A. 2019. A Stress-responsive signaling network regulating pseudohyphal growth and ribonucleoprotein granule abundance in Saccharomyces cerevisiae. Genetics 213, 705–720.

    Article  CAS  Google Scholar 

  • Park, Y.U., Hur, H., Ka, M., and Kim, J. 2006. Identification of translational regulation target genes during filamentous growth in Saccharomyces cerevisiae: regulatory role of Caf20 and Dhh1. Eukaryot. Cell 5, 2120–2127.

    Article  CAS  Google Scholar 

  • Reijns, M.A.M., Alexander, R.D., Spiller, M.P., and Beggs, J.D. 2008. A role for Q/N-rich aggregation-prone regions in P-body localization. J. Cell Sci. 121, 2463–2472.

    Article  CAS  Google Scholar 

  • Roberts, R.L. and Fink, G.R. 1994. Elements of a single MAP kinase cascade in Saccharomyces cerevisiae mediate two developmental programs in the same cell type: mating and invasive growth. Genes Dev. 8, 2974–2985.

    Article  CAS  Google Scholar 

  • Shively, C.A., Kweon, H.K., Norman, K.L., Mellacheruvu, D., Xu, T., Sheidy, D.T., Dobry, C.J., Sabath, I., Cosky, E.E.P., Tran, E.J., et al. 2015. Large-scale analysis of kinase signaling in yeast pseudohyphal development identifies regulation of ribonucleoprotein granules. PLoS Genet. 11, e1005564.

    Article  Google Scholar 

  • Sikorski, R.S. and Hieter, P. 1989. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122, 19–27.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sudbery, P., Gow, N., and Berman, J. 2004. The distinct morphogenic states of Candida albicans. Trends Microbiol. 12, 317–324.

    Article  CAS  Google Scholar 

  • Sweet, T., Kovalak, C., and Coller, J. 2012. The DEAD-box protein Dhh1 promotes decapping by slowing ribosome movement. PLoS Biol. 10, e1001342.

    Article  CAS  Google Scholar 

  • Tanner, N.K., Cordin, O., Banroques, J., Doere, M., and Linder, P. 2003. The Q motif: a newly identified motif in DEAD box helicases may regulate ATP binding and hydrolysis. Mol. Cell 11, 127–138.

    Article  CAS  Google Scholar 

  • Teixeira, D. and Parker, R. 2007. Analysis of P-body assembly in Saccharomyces cerevisiae. Mol. Biol. Cell 18, 2274–2287.

    Article  CAS  Google Scholar 

  • Zhang, Q., Meng, X., Li, D., Chen, S., Luo, J., Zhu, L., Singer, R.H., and Gu, W. 2017. Binding of DEAD-box helicase Dhh1 to the 5'-untranslated region of ASH1 mRNA represses localized translation of ASH1 in yeast cells. J. Biol. Chem. 292, 9787–9800.

    Article  CAS  Google Scholar 

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Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (2015R1C1A2A01051577).

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Correspondence to Jinmi Kim.

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Lee, E., Jung, D. & Kim, J. Roles of Dhh1 RNA helicase in yeast filamentous growth: Analysis of N-terminal phosphorylation residues and ATPase domains. J Microbiol. 58, 853–858 (2020). https://doi.org/10.1007/s12275-020-0431-7

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  • DOI: https://doi.org/10.1007/s12275-020-0431-7

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