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Current Genetics

, Volume 65, Issue 5, pp 1251–1261 | Cite as

Identification and functional characterization of Candida albicans mannose–ethanolamine phosphotransferase (Mcd4p)

  • Satoru Hasegawa
  • Yuimi Yamada
  • Noboru Iwanami
  • Yusuke Nakayama
  • Hironobu Nakayama
  • Shun Iwatani
  • Takahiro Oura
  • Susumu KajiwaraEmail author
Original Article
  • 101 Downloads

Abstract

Glycosylphosphatidylinositol (GPI) is an important compound for the growth of fungi, because GPI-anchored proteins including glycosyltransferases and adhesins are involved in cell-wall integrity, adhesion, and nutrient uptake in this organism. In this study, we examined orf19.5244 in the genome database of the pathogenic fungus Candida albicans, a homologue of the Saccharomyces cerevisiae mannose–ethanolamine phosphotransferase gene, MCD4, which plays a role in GPI synthesis. Expression of this homologue, designated CaMCD4, restored cell growth in a defective conditional mcd4 mutant of S. cerevisiae, Scmcd4t, in which expression of native MCD4 was repressed in the presence of doxycycline (Dox). Analysis of radiolabeled lipids showed that the accumulation of abnormal GPI anchor precursors in Scmcd4t decreased markedly upon expression of CaMCD4. Moreover, we constructed a single mutant (Camcd4/CaMCD4) and a conditional double mutant (Camcd4/Camcd4t) at the MCD4 locus of C. albicans. Repression of CaMCD4 expression by Dox led to a decrease in growth and appearance of abnormal morphology in C. albicans, both in vitro and in a silkworm infection model. These results suggest that CaMcd4p is indispensable for growth of C. albicans both in vitro and in infected hosts and a candidate target for the development of new antifungals.

Keywords

Glycosylphosphatidylinositol (GPI)-anchor synthesis Candida albicans Codon optimization 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The procedures performed in studies involving animals were in accordance with the ethical standards of the institution and this article does not contain any studies with participants performed by any of the authors.

Supplementary material

294_2019_987_MOESM1_ESM.pptx (65 kb)
Supplemented Figure 1 Southern blot analysis of C. albicans transformants. The BglII-digested genome DNAs from TUA4, Camcd4::ARG4/CaMCD4, and Camcd4::ARG4/Camcd4::URA3 strains were used. The 0.5 kb DNA fragment corresponding to the 5’ end of CaMCD4 was amplified by PCR and used as a radiolabeled probe. The hybridization conditions were shown in “Methods”. Table 1 Strains used in this study. (PPTX 64 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Life Science and TechnologyTokyo Institute of TechnologyYokohamaJapan
  2. 2.Faculty of Pharmaceutical SciencesSuzuka University of Medical ScienceSuzukaJapan

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