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Cryptococcus neoformans sexual reproduction is controlled by a quorum sensing peptide

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Abstract

Bacterial quorum sensing is a well-characterized communication system that governs a large variety of collective behaviours. By comparison, quorum sensing regulation in eukaryotic microbes remains poorly understood, especially its functional role in eukaryote-specific behaviours, such as sexual reproduction. Cryptococcus neoformans is a prevalent fungal pathogen that has two defined sexual cycles (bisexual and unisexual) and is a model organism for studying sexual reproduction in fungi. Here, we show that the quorum sensing peptide Qsp1 serves as an important signalling molecule for both forms of sexual reproduction. Qsp1 orchestrates various differentiation and molecular processes, including meiosis, the hallmark of sexual reproduction. It activates bisexual mating, at least in part through the control of pheromone, a signal necessary for bisexual activation. Notably, Qsp1 also plays a major role in the intercellular regulation of unisexual initiation and coordination, in which pheromone is not strictly required. Through a multi-layered genetic screening approach, we identified the atypical zinc finger regulator Cqs2 as an important component of the Qsp1 signalling cascade during both bisexual and unisexual reproduction. The absence of Cqs2 eliminates the Qsp1-stimulated mating response. Together, these findings extend the range of behaviours governed by quorum sensing to sexual development and meiosis.

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Fig. 1: The mating pheromone is largely dispensable for unisexual reproduction in the XL280α background.
Fig. 2: Quorum sensing peptide encoding gene CQS1 is important for both bisexual and unisexual development.
Fig. 3: Qsp1 mediates mating response through the control of Mat2.
Fig. 4: Qsp1 participates in glucose starvation-induced mating response and morphogenesis.
Fig. 5: Cqs2 as an important Qsp1 signalling cascade member promotes bisexual and unisexual mating.
Fig. 6: Cqs2 can associate with its own promoter through an atypical C2H2 zinc finger domain conserved among divergent fungal species.

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Acknowledgements

We thank P. Sharma and Wang laboratory members for critical reading, and S. Luo and H. Liu for their assistance in genetic screening. This work was financially supported by the National Science and Technology Major Project (2018ZX10101004), the Key Research Program of the Chinese Academy of Sciences (QYZDB-SSW-SSMC040), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDPB03), the National Natural Science Foundation of China (Grants 31622004, 31570138, 31501008 and 31501009) and the National Institutes of Health (http://www.niaid.nih.gov/Pages/default.aspx) (R01AI097599 to X.L.). L.W. is a member of the Thousand Talents Program. X.L. holds an Investigator Award in the Pathogenesis of Infectious Disease from the Burroughs Wellcome Fund (http://www.bwfund.org/).

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Author notes

  1. These authors contributed equally to this work: Xiuyun Tian, Guang-Jun He, Pengjie Hu.

Authors and Affiliations

  1. State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China

    Xiuyun Tian, Guang-Jun He, Pengjie Hu, Lei Chen, Lan Shen, Weixin Ke, Haijiao Xu, Fengyan Bai & Linqi Wang

  2. University of Chinese Academy of Sciences, Beijing, China

    Pengjie Hu, Lei Chen, Weixin Ke & Linqi Wang

  3. Department of Microbiology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China

    Changyu Tao & Ence Yang

  4. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China

    Ying-Lu Cui & Bian Wu

  5. Department of Microbiology, University of Georgia, Athens, GA, USA

    Youbao Zhao & Xiaorong Lin

  6. College of Life Science, Northeast Forestry University, Harbin, China

    Qijiang Xu

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Contributions

All authors contributed to the data analysis. X.T., G.-J.H., P.H., L.C., C.T., Y.-L.C., L.S., W.K., H.X., F.B., B.W., E.Y., X.L. and L.W. designed the experiments. X.T., G.-J.H. and P.H. conducted most of the studies. L.C. and E.Y. conducted most of the bioinformatics assays. Y.-L.C. and B.W. conducted the Rosetta ab initio and comparative modelling. L.S. conducted part of the overexpression plasmids construction. H.X. conducted the subcellular localization analysis. Y.Z. conducted part of the gene deletion experiments. Q.X., F.B., B.W., E.Y., X.L. and L.W. contributed reagents/materials/analysis tools. X.T., G.-J.H., X.L. and L.W. wrote the paper with contributions from all other authors.

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Correspondence to Linqi Wang.

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Supplementary information

Supplementary Information

Supplementary Figures 1–9, Supplementary Table 3.

Reporting Summary

Supplementary Table 1

Cryptococcus neoformans XL280α gene expression in response to different signals (mating cue, Qsp1, Qsp2 and Qsp3 peptides).

Supplementary Table 2

Expression profiles of the wild-type strain (XL280α) and the cqs1Δ mutant in the presence of glucose or galactose.

Supplementary Table 4

Genes differentially expressed in the opt1Δ mutant during unisexual mating.

Supplementary Table 5

Genes differentially expressed in the cqs2Δ mutant in the presence or absence of Qsp1 under the mating-inducing condition.

Supplementary Table 6

Strains used in this study.

Supplementary Table 7

Primers used in this study.

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Tian, X., He, GJ., Hu, P. et al. Cryptococcus neoformans sexual reproduction is controlled by a quorum sensing peptide. Nat Microbiol 3, 698–707 (2018). https://doi.org/10.1038/s41564-018-0160-4

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