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Identifying Leptospira interrogans putative virulence factors with a yeast protein expression screen

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Abstract

Leptospirosis is a zoonotic disease caused by pathogenic Leptospira spp., with global implications primarily in tropical countries. However, the mechanisms of leptospiral pathogenesis are still not fully known and not all virulence factors (VFs) have been identified. Budding yeast, Saccharomyces cerevisiae is a popular eukaryotic model which has been used to identify bacterial VFs that target the conserved eukaryotic cellular processes. In this study, we screened for putative VFs of L. interrogans, one of the dominant species causing leptospirosis, by expressing candidate VFs in budding yeast and examining their impact on yeast growth in a high-throughput format. From an initial selection of 288 L. interrogans ORFs, we screened 226 candidate VFs in a yeast growth inhibition assay and identified nine putative VFs in four categories (adhesion, enzymatic, host structure interaction, and immunogenicity). Notably, LIC10280 was highly toxic even when expressed at low copies. We also observed specific subcellular localization for several putative VFs. This study shows that there are still potential L. interrogans VFs that await discovery.

Key points

• High-throughput cloning and expression of leptospiral proteins in yeast.

• Heterologous expression of nine leptospiral proteins inhibited yeast growth.

• An uncharacterized protein LIC10280 maybe a putative VF for further validation.

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Data availability

All data generated or analyzed during this study are included in this published article [and its supplementary information files].

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Acknowledgements

The authors would like to thank Professor Yoshikazu Ohya from the Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, Japan, for training in yeast staining, and the Bioprobe Application Research Unit, RIKEN Centre for Sustainable Resource Science, for the use of the Bio Microplate Reader HiTS (Cosmo Bio, Japan).

Funding

This study was funded by the USM RUI grants number 1001/CIPPM/812147 and 1001/CIPPM/8012262 awarded to EBBO, MRS, AT, and RN.

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Contributions

WYL and ZW carried out experiments and data analysis. WYL and ZW drafted the manuscript. EBBO conceived the idea, designed the experiments, and analyzed the data. All authors were involved in discussions, review, and manuscript revision.

Corresponding author

Correspondence to Eugene Boon Beng Ong.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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All authors declare no competing interests.

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Lai, W.Y., Wong, Z., Chang, C.H. et al. Identifying Leptospira interrogans putative virulence factors with a yeast protein expression screen. Appl Microbiol Biotechnol 106, 6567–6581 (2022). https://doi.org/10.1007/s00253-022-12160-1

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  • DOI: https://doi.org/10.1007/s00253-022-12160-1

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