Reporters for the analysis of N-glycosylation in Candida albicans

Shahida Shahana, Hector M Mora-Montes, Luis Castillo, Iryna Bohovych, Chirag C Sheth, Frank C Odds, Neil A R Gow, Alistair J P Brown

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Abstract

A large proportion of Candida albicans cell surface proteins are decorated post-translationally by glycosylation. Indeed N-glycosylation is critical for cell wall biogenesis in this major fungal pathogen and for its interactions with host cells. A detailed understanding of N-glycosylation will yield deeper insights into host-pathogen interactions. However, the analysis of N-glycosylation is extremely challenging because of the complexity and heterogeneity of these structures. Therefore, in an attempt to reduce this complexity and facilitate the analysis of N-glycosylation, we have developed new synthetic C. albicans reporters that carry a single N-linked glycosylation site derived from Saccharomyces cerevisiae Suc2. These glycosylation reporters, which carry C.albicans Hex1 or Sap2 signal sequences plus carboxy-terminal FLAG3 and His6 tags, were expressed in C.albicans from the ACT1 promoter. The reporter proteins were successfully secreted and hyperglycosylated by C.albicans cells, and their outer chain glycosylation was dependent on Och1 and Pmr1, which are required for N-mannan synthesis, but not on Mnt1 and Mnt2 which are only required for O-mannosylation. These reporters are useful tools for the experimental dissection of N-glycosylation and other related processes in C.albicans, such as secretion.
Original languageEnglish
Pages (from-to)107-115
Number of pages9
JournalFungal Genetics and Biology
Volume56
Early online date19 Apr 2013
DOIs
Publication statusPublished - Jul 2013

Keywords

  • candida albicans
  • glycosylation
  • cell wall
  • glycosylation reporter

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    Shahana, S., Mora-Montes, H. M., Castillo, L., Bohovych, I., Sheth, C. C., Odds, F. C., Gow, N. A. R., & Brown, A. J. P. (2013). Reporters for the analysis of N-glycosylation in Candida albicans. Fungal Genetics and Biology, 56, 107-115. https://doi.org/10.1016/j.fgb.2013.03.009