TY  - JOUR
A1  - Götz, Ralph
A1  - Panzer, Sabine
A1  - Trinks, Nora
A1  - Eilts, Janna
A1  - Wagener, Johannes
A1  - Turrà, David
A1  - Di Pietro, Antonio
A1  - Sauer, Markus
A1  - Terpitz, Ulrich
T1  - Expansion Microscopy for Cell Biology Analysis in Fungi
JF  - Frontiers in Microbiology
N2  - Super-resolution microscopy has evolved as a powerful method for subdiffraction-resolution fluorescence imaging of cells and cellular organelles, but requires sophisticated and expensive installations. Expansion microscopy (ExM), which is based on the physical expansion of the cellular structure of interest, provides a cheap alternative to bypass the diffraction limit and enable super-resolution imaging on a conventional fluorescence microscope. While ExM has shown impressive results for the magnified visualization of proteins and RNAs in cells and tissues, it has not yet been applied in fungi, mainly due to their complex cell wall. Here we developed a method that enables reliable isotropic expansion of ascomycetes and basidiomycetes upon treatment with cell wall degrading enzymes. Confocal laser scanning microscopy (CLSM) and structured illumination microscopy (SIM) images of 4.5-fold expanded sporidia of Ustilago maydis expressing fluorescent fungal rhodopsins and hyphae of Fusarium oxysporum or Aspergillus fumigatus expressing either histone H1-mCherry together with Lifeact-sGFP or mRFP targeted to mitochondria, revealed details of subcellular structures with an estimated spatial resolution of around 30 nm. ExM is thus well suited for cell biology studies in fungi on conventional fluorescence microscopes.
KW  - Expansion microscopy
KW  - fluorescence microscopy
KW  - fungi
KW  - sporidia
KW  - hyphae
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-202569
SN  - 1664-302X
VL  - 11
ER  - 
TY  - JOUR
A1  - Bar-Yosef, Hagit
A1  - Gildor, Tsvia
A1  - Ramírez-Zavala, Bernardo
A1  - Schmauch, Christian
A1  - Weissman, Ziva
A1  - Pinsky, Mariel
A1  - Naddaf, Rawi
A1  - Morschhäuser, Joachim
A1  - Arkowitz, Robert A.
A1  - Kornitzer, Daniel
T1  - A global analysis of kinase function in Candida albicans hyphal morphogenesis reveals a role for the endocytosis regulator Akl1
JF  - Frontiers in Cellular and Infection Microbiology
N2  - The human pathogenic fungus Candida albicans can switch between yeast and hyphal morphologies as a function of environmental conditions and cellular physiology. The yeast-to-hyphae morphogenetic switch is activated by well-established, kinase-based signal transduction pathways that are induced by extracellular stimuli. In order to identify possible inhibitory pathways of the yeast-to-hyphae transition, we interrogated a collection of C. albicans protein kinases and phosphatases ectopically expressed under the regulation of the TETon promoter. Proportionately more phosphatases than kinases were identified that inhibited hyphal morphogenesis, consistent with the known role of protein phosphorylation in hyphal induction. Among the kinases, we identified AKL1 as a gene that significantly suppressed hyphal morphogenesis in serum. Akl1 specifically affected hyphal elongation rather than initiation: overexpression of AKL1 repressed hyphal growth, and deletion of AKL1 resulted in acceleration of the rate of hyphal elongation. Akl1 suppressed fluid-phase endocytosis, probably via Pan1, a putative clathrin-mediated endocytosis scaffolding protein. In the absence of Akl1, the Pan1 patches were delocalized from the sub-apical region, and fluid-phase endocytosis was intensified. These results underscore the requirement of an active endocytic pathway for hyphal morphogenesis. Furthermore, these results suggest that under standard conditions, endocytosis is rate-limiting for hyphal elongation.
KW  - hyphae
KW  - endocytosis
KW  - Pan1
KW  - functional genomics
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-197204
SN  - 2235-2988
VL  - 8
ER  -