Immunolocalization of chitin synthases in the phytopathogenic dimorphic fungus Ustilago maydis

Conserved polypeptides of the chitin synthase genes UmCHS3 and UmCHS6 from the phytopathogenic fungus Ustilago maydis were utilized as immunogens to obtain polyclonal antibodies that were purified by affinity procedures. Because of their similarities at the regions encoded by either polypeptide, it...

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Bibliographic Details
Published in:FEMS yeast research 2006-11, Vol.6 (7), p.999-1009
Main Authors: Ruiz-Herrera, José, Xoconostle-Cázares, Beatriz, Reynaga-Peña, Cristina G, León-Ramírez, Claudia, Cárabez-Trejo, Alfonso
Format: Article
Language:English
Subjects:
cell wall
Centrifugation, Density Gradient
Chitin
Chitin synthase
Chitin Synthase - analysis
chitosomes
Cytoplasm
Electron microscopy
Immunoblotting
Immunoelectron microscopy
Immunofluorescence
Immunoglobulins
Immunohistochemistry
Localization
Microscopy
Microscopy, Electron
Microscopy, Fluorescence
Phytopathogenic fungi
Polyclonal antibodies
Polypeptides
Ustilago - enzymology
Ustilago maydis
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Summary:Conserved polypeptides of the chitin synthase genes UmCHS3 and UmCHS6 from the phytopathogenic fungus Ustilago maydis were utilized as immunogens to obtain polyclonal antibodies that were purified by affinity procedures. Because of their similarities at the regions encoded by either polypeptide, it was concluded that anti-Chs3 antibodies recognized both Chs3 and Chs4 chitin synthases, whereas anti-Chs6 antibodies recognized Chs6 and Chs8 polypeptides. These antibodies were used to analyze the localization of the corresponding chitin synthases in U. maydis cells, using both indirect immunofluorescence microscopy and immunoelectron microscopy with colloidal-gold-labeled secondary antibodies. It was observed that chitin synthase proteins were accumulated both in the surface and in the cytoplasm of the fungal cells. Electron microscopy images revealed the accumulation of clusters of gold particles in vesicles, providing evidence for the possible origin and destination of chitin synthases in the fungal cells.
ISSN:1567-1356
1567-1364
DOI:10.1111/j.1567-1364.2006.00133.x