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Co-delivery of cell-wall-forming enzymes in the same vesicle for coordinated fungal cell wall formation

Nature Microbiology volume 1, Article number: 16149 (2016) Cite this article

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Abstract

Fungal cells are surrounded by an extracellular cell wall. This complex matrix of proteins and polysaccharides protects against adverse stresses and determines the shape of fungal cells. The polysaccharides of the fungal wall include 1,3-β-glucan and chitin, which are synthesized by membrane-bound synthases at the growing cell tip. A hallmark of filamentous fungi is the class V chitin synthase, which carries a myosin-motor domain. In the corn smut fungus Ustilago maydis, the myosin-chitin synthase Mcs1 moves to the plasma membrane in secretory vesicles, being delivered by kinesin-1 and myosin-5. The myosin domain of Mcs1 enhances polar secretion by tethering vesicles at the site of exocytosis. It remains elusive, however, how other cell-wall-forming enzymes are delivered and how their activity is coordinated post secretion. Here, we show that the U. maydis class VII chitin synthase and 1,3-β-glucan synthase travel in Mcs1-containing vesicles, and that their apical secretion depends on Mcs1. Once in the plasma membrane, anchorage requires enzyme activity, which suggests co-synthesis of chitin and 1,3-β-glucan polysaccharides at sites of exocytosis. Thus, delivery of cell-wall-forming enzymes in Mcs1 vesicles ensures local foci of fungal cell wall formation.

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Figure 1: Domain organization of fungal class V and class VII CHSs.
Figure 2: Class V CHS and class VII CHS co-migrate and are co-secreted.
Figure 3: Null mutants of mcs1 and chs6 exhibit different cell wall phenotypes.
Figure 4: Mcs1, Chs6 and Gsc1 locate in the same vesicle.
Figure 5: Mcs1, Chs6 and Gsc1 remain co-located in stationary wall-forming foci.
Figure 6: Cell wall synthases co-travel in the same vesicle in hyphae.

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Acknowledgements

The authors thank U. Fuchs, S. Milne and P. Splatt for technical support. M.M.-U. thanks N.J. Talbot for financial support. G.S. acknowledges J. Stajich for fruitful discussions. This work was supported by the Biotechnology & Biological Sciences Research Council (grants BB/H019774/1 and BB/I020667/1 to G.S.).

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Author notes

  1. Martin Schuster & Magdalena Martin-Urdiroz

    Present address: †Present address: Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1, Fukuoka 812-8581, Japan,

  2. Yujiro Higuchi: These authors contributed equally to this work.

Authors and Affiliations

  1. Biosciences, University of Exeter, Stocker Road, Exeter EX4 4QD, UK

    Martin Schuster, Magdalena Martin-Urdiroz, Yujiro Higuchi, Christian Hacker, Sreedhar Kilaru, Sarah J. Gurr & Gero Steinberg

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Contributions

G.S. developed the research plan and the experimental strategy, directed the project, analysed data, assembled all figures and videos and wrote the manuscript (with the exception of the Methods, which was written mainly by M.M.-U., M.S. and C.H.). M.M.-U. generated strains and plasmids, performed experiments and analysed data. M.S. performed experiments, generated strains and analysed data. Y.H. and S.K. generated strains and plasmids. C.H. performed electron microscopy. S.J.G. discussed the project and data. All authors corrected the manuscript and discussed data.

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Correspondence to Gero Steinberg.

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Supplementary information

Supplementary Information

Supplementary Figures 1-6 (Colour-adjusted main figures for readers with red-green colour blindness), Supplementary Figures 7-16, Supplementary Tables 1-3, Supplementary References (PDF 3612 kb)

Supplementary Video 1

Motility of Chs6-GFP3 in U. Maydis. (MOV 429 kb)

Supplementary Video 2

Motility of Mcs1-GFP3 and Chs6-GFP3 in a yeast-like cell of U. maydis. (MOV 417 kb)

Supplementary Video 3

Co-motility of mCherry3-Mcs1 and Chs6-GFP3 in a yeast-like cell of U. maydis. (MOV 127 kb)

Supplementary Video 4

Co-motility of mCherry3-Mcs1 and GFP3-Gsc1 in a yeast-like cell of U. maydis. (MOV 226 kb)

Supplementary Video 5

Diffusive motion of Mcs1-GFP3 in control cells, cell wall-less protoplasts and in the presence of the CHS inhibitor nikkomycin Z and the GS inhibitor caspofungin. (MOV 448 kb)

Supplementary Video 6

Co-motility of mCherry3-Mcs1 and GFP3-Gsc1 in a hyphal cell of U. maydis. (MOV 251 kb)

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Schuster, M., Martin-Urdiroz, M., Higuchi, Y. et al. Co-delivery of cell-wall-forming enzymes in the same vesicle for coordinated fungal cell wall formation. Nat Microbiol 1, 16149 (2016). https://doi.org/10.1038/nmicrobiol.2016.149

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