title: Functional characterization of extracellular chitinase encoded by the YlCTS1 gene in a dimorphic yeast Yarrowia lipolytica.
abstract: The hemiascomycetes yeast Yarrowia lipolytica is a dimorphic yeast with alternating yeast and mycelia forms. Bioinformatic analysis revealed the presence of three putative chitinase genes, YlCTS1, YlCTS2, and YlCTS3, in the Y. lipolytica genome. Here, we demonstrated that the protein of YlCTS1 (YlCts1p), which contains an N-terminal secretion signal peptide, a long C-terminal Ser/Thr-rich domain, and a chitin-binding domain, is a homologue to Saccharomyces cerevisiae chitinase 1 (ScCts1p). Deletion of YlCTS1 remarkably reduced extracellular endochitinase activity in the culture supernatant of Y. lipolytica and enhanced cell aggregation, suggesting a role of YlCts1p in cell separation as ScCts1p does in S. cerevisiae. However, loss of YlCts1p function did not affect hyphal formation induced by fetal bovine serum addition. The mass of YlCts1p was dramatically decreased by jack bean α-mannosidase digestion but not by PNGase F treatment, indicating that YlCts1p is modified only by O-mannosylation without N-glycosylation. Moreover, the O-glycan profile of YlCts1p was identical to that of total cell wall mannoproteins, supporting the notion that YlCts1p can be used as a good model for studying O-glycosylation in this dimorphic yeast.
authors.affiliation: Department of Life Science, Chung-Ang University, Seoul, 156-756, Republic of Korea.
authors.forename: Jeong-Nam
authors.initials: JN
authors.lastname: Park
authors.forename: Chang Pyo
authors.initials: CP
authors.lastname: Han
authors.forename: Dong-Jik
authors.initials: DJ
authors.lastname: Lee
authors.forename: Seon Ah
authors.initials: SA
authors.lastname: Cheon
authors.forename: Hyun Ah
authors.initials: HA
authors.lastname: Kang
mesh_terms: D002448:Cell Adhesion; D002688:Chitinases / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D005800:Genes, Fungal; D006031:Glycosylation; D008358:Mannose / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics
chemical_list: D029701:Saccharomyces cerevisiae Proteins; C121990:CTS1 protein, S cerevisiae; D002688:Chitinases; D008358:Mannose