Traffic of Chitin Synthase 1 (CHS-1) to the Spitzenkörper and Developing Septa in Hyphae of Neurospora crassa: Actin Dependence and Evidence of Distinct Microvesicle Populations
| Autor: | Salomon Bartnicki-Garcia, Robert W. Roberson, Eddy Sánchez-León, Meritxell Riquelme, Michael Freitag, Jorge Verdín |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Green Fluorescent Proteins
Hyphae Biology Microbiology Neurospora crassa Fungal Proteins hemic and lymphatic diseases Fluorescence microscope skin and connective tissue diseases Molecular Biology Chitin Synthase Fungal protein integumentary system Microvesicle Cytoplasmic Vesicles fungi Spitzenkörper Articles General Medicine Chitin synthase Actin cytoskeleton biology.organism_classification Actins Cell biology Actin Cytoskeleton Microscopy Fluorescence Biochemistry Cytoplasm biology.protein |
| Zdroj: | Eukaryotic Cell. 10:683-695 |
| ISSN: | 1535-9786 1535-9778 |
| DOI: | 10.1128/ec.00280-10 |
| Popis: | We describe the subcellular location of chitin synthase 1 (CHS-1), one of seven chitin synthases in Neurospora crassa . Laser scanning confocal microscopy of growing hyphae showed CHS-1–green fluorescent protein (GFP) localized conspicuously in regions of active wall synthesis, namely, the core of the Spitzenkörper (Spk), the apical cell surface, and developing septa. It was also present in numerous fine particles throughout the cytoplasm plus some large vacuoles in distal hyphal regions. Although the same general subcellular distribution was observed previously for CHS-3 and CHS-6, they did not fully colocalize. Dual labeling showed that the three different chitin synthases were contained in different vesicular compartments, suggesting the existence of a different subpopulation of chitosomes for each CHS. CHS-1–GFP persisted in the Spk during hyphal elongation but disappeared from the septum after its development was completed. Wide-field fluorescence microscopy and total internal reflection fluorescence microscopy revealed subapical clouds of particles, suggestive of chitosomes moving continuously toward the Spk. Benomyl had no effect on CHS-1–GFP localization, indicating that microtubules are not strictly required for CHS trafficking to the hyphal apex. Conversely, actin inhibitors caused severe mislocalization of CHS-1–GFP, indicating that actin plays a major role in the orderly traffic and localization of CHS-1 at the apex. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|