title: Scanning and negative-staining electron microscopy of protoplast regeneration of a wild-type and two chitin synthase mutants in the pathogenic yeast Candida glabrata.
abstract: Protoplast regeneration of a wild-type and two mutant strains of Candida glabrata defective in CHS3 homologues encoding class IV chitin synthase in Saccharomyces cerevisiae was examined by scanning and negative-staining electron microscopy. In the wild-type strain, small particles and short filaments appeared on the protoplast surface at 10 min, filamentous materials covered the entire surface of the protoplast at 1 h, granular materials started filling interspaces of filamentous materials at 2 h and regeneration was completed at 6 h. The filamentous materials consisted of microfibrils of various widths ranging from ≤5 to 40 nm, and composed of β-glucan. Protoplasts of the two chitin synthase mutant strains of Δchs3A and Δchs3B completed regeneration essentially by the same process as wild-type strain, although it took more time. These results suggest that CHS3A and CHS3B genes may have important roles in cell wall formation during protoplast regeneration, but can be compensated by other cell wall enzymes.
authors.affiliation: Medical Mycology Research Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8673, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Namiki
authors.forename: Keigo
authors.initials: K
authors.lastname: Ueno
authors.forename: Hiroki
authors.initials: H
authors.lastname: Mitani
authors.forename: Eric V
authors.initials: EV
authors.lastname: Virtudazo
authors.forename: Misako
authors.initials: M
authors.lastname: Ohkusu
authors.forename: Kiminori
authors.initials: K
authors.lastname: Shimizu
authors.forename: Susumu
authors.initials: S
authors.lastname: Kawamoto
authors.forename: Hiroji
authors.initials: H
authors.lastname: Chibana
authors.forename: Masashi
authors.initials: M
authors.lastname: Yamaguchi
mesh_terms: D041221:Candida glabrata / Q000201:enzymology / Q000472:pathogenicity / Q000502:physiology / Q000648:ultrastructure*; D002473:Cell Wall / Q000378:metabolism* / Q000648:ultrastructure; D002687:Chitin Synthase / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005936:Glucans / Q000648:ultrastructure; D020894:Microfibrils / Q000648:ultrastructure; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D009154:Mutation; D016624:Negative Staining; D011523:Protoplasts / Q000502:physiology* / Q000648:ultrastructure*
chemical_list: D005656:Fungal Proteins; D005936:Glucans; D002687:Chitin Synthase