PilY1 and minor pilins form a complex priming the type IVa pilus in Myxococcus xanthus
Abstract
Type IVa pili are ubiquitous and versatile bacterial cell surface filaments that undergo cycles of extension, adhesion and retraction powered by the cell-envelope spanning type IVa pilus machine (T4aPM). The overall architecture of the T4aPM and the location of 10 conserved core proteins within this architecture have been elucidated. Here, using genetics, cell biology, proteomics and cryo-electron tomography, we demonstrate that the PilY1 protein and four minor pilins, which are widely conserved in T4aP systems, are essential for pilus extension in Myxococcus xanthus and form a complex that is an integral part of the T4aPM. Moreover, these proteins are part of the extended pilus. Our data support a model whereby the PilY1/minor pilin complex functions as a priming complex in T4aPM for pilus extension, a tip complex in the extended pilus for adhesion, and a cork for terminating retraction to maintain a priming complex for the next round of extension.
Additional Information
© The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 30 September 2019. Accepted 14 September 2020. Published 07 October 2020. We thank Ahmet Tekin for strains and plasmids and Magdalena Anna Świątek-Połatyńska for LonD antibodies. Cryo-ET work was done in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech. This work was supported by the Deutsche Forschungsgemeinschaft (DFG) within the framework of the SFB987 "Microbial Diversity in Environmental Signal response" (to L.S.-A.), the Max Planck Society (to L.S.-A.), and NIH grant RO1 AI27401 (to G.J.J.). Data availability. The proteomics datasets are available via the PRIDE database (ProteomeXchange accession: PXD021163). We also used the following publicly available datasets: Uniprot (https://www.uniprot.org/) and KEGG (https://www.genome.jp/kegg/), including the KEGG SSDB database (https://www.kegg.jp/kegg/ssdb/). Source data are provided with this paper. Open Access funding enabled and organized by Projekt DEAL. Author Contributions. A.T.-L.: designed and conceived the study and performed most of the experiments. Y.-W.C.: performed the cryo-electron tomographic experiments and the modeling of the priming complex. T.G.: performed purification and analysis of pull-down samples and label-free mass spectrometry-based quantitative proteomics. S.L.: generated plasmids and strains and performed motility assays. M.H.: generated plasmids and strains and helped with transmission electron microscopy and bioinformatics. G.C.: helped with cryo-electron tomography data acquisition. G.J.J. and L.S.-A.: conceived the study, supervised research and provided funding. A.T.-L., Y.-W.C., T.G., G.J.J., and L.S.-A.: analyzed and interpreted data and wrote the manuscript. All authors approved the final manuscript. The authors declare no competing interests. Reporting summary. Further information on research design is available in the Nature Research Reporting Summary linked to this article. Peer review information. Nature Communications thanks Lisa Craig and the other anonymous reviewers for their contribution to the peer review of this work. Peer reviewer reports are available.Attached Files
Published - s41467-020-18803-z.pdf
Supplemental Material - 41467_2020_18803_MOESM1_ESM.pdf
Supplemental Material - 41467_2020_18803_MOESM2_ESM.pdf
Supplemental Material - 41467_2020_18803_MOESM3_ESM.pdf
Supplemental Material - 41467_2020_18803_MOESM4_ESM.xlsx
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Additional details
- PMCID
- PMC7541494
- Eprint ID
- 105912
- Resolver ID
- CaltechAUTHORS:20201008-083808921
- SFB987
- Deutsche Forschungsgemeinschaft (DFG)
- Max-Planck-Gesellschaft
- R01 AI27401
- NIH
- Created
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2020-10-08Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering