Τίτλος:
Superresolution and pulse-chase imaging reveal the role of vesicle transport in polar growth of fungal cells
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
Polarized growth of filamentous fungi requires continuous transport of biomolecules to the hyphal tip. To this end, construction materials are packaged in vesicles and transported by motor proteins along microtubules and actin filaments. We have studied these processes with quantitative superresolution localization microscopy of live Aspergillus nidulans cells expressing the photoconvertible protein mEosFPthermo fused to the chitin synthase ChsB. ChsB is mainly located at the Spitzenkörper near the hyphal tip and produces chitin, a key component of the cell wall. We have visualized the pulsatory dynamics of the Spitzenkörper, reflecting vesicle accumulation before exocytosis and their subsequent fusion with the apical plasma membrane. Furthermore, high-speed pulse-chase imaging after photoconversion of mEosFPthermo in a tightly focused spot revealed that ChsB is transported with two different speeds from the cell body to the hyphal tip and vice versa. Comparative analysis using motor protein deletion mutants allowed us to assign the fast movements (7 to 10 mm s−1) to transport of secretory vesicles by kinesin-1, and the slower ones (2 to 7 mm s−1) to transport by kinesin-3 on early endosomes. Our results show how motor proteins ensure the supply of vesicles to the hyphal tip, where temporally regulated exocytosis results in stepwise tip extension. Copyright © 2018 The Authors.
Συγγραφείς:
Zhou, L.
Evangelinos, M.
Wernet, V.
Eckert, A.F.
Ishitsuka, Y.
Fischer, R.
Nienhaus, G.U.
Takeshita, N.
Περιοδικό:
Science Advances
Εκδότης:
American Association for the Advancement of Science
Λέξεις-κλειδιά:
Cell membranes; Cells; Chitin; Cytology; Optical resolving power, Apical plasma membrane; Aspergillus nidulans; Comparative analysis; Deletion mutants; Filamentous fungi; Secretory vesicles; Super resolution; Vesicle transport, Proteins, chitin synthase; fungal protein, actin filament; Aspergillus nidulans; cytology; fungus hyphae; genetics; growth, development and aging; light; metabolism; microtubule; mutation; physiology; three dimensional imaging; transport vesicle, Actin Cytoskeleton; Aspergillus nidulans; Chitin Synthase; Fungal Proteins; Hyphae; Imaging, Three-Dimensional; Light; Microtubules; Mutation; Transport Vesicles
DOI:
10.1126/sciadv.1701798
