Analysis of extracellular vesicles produced in the biofilm by the dimorphic yeast Pichia fermentans

J Cell Physiol. 2018 Apr;233(4):2759-2767. doi: 10.1002/jcp.25885. Epub 2017 May 16.

Abstract

The yeast Pichia fermentans DISAABA 726 strain (P. fermentans) is a dimorphic yeast that under different environmental conditions may switch from a yeast-like to pseudohyphal morphology. We hypothesize that exosomes-like vesicles (EV) could mediate this rapid modification. EV are membrane-derived vesicles carrying lipids, proteins, mRNAs and microRNAs and have been recognized as important mediators of intercellular communication. Although it has been assumed for a long time that fungi release EV, knowledge of their functions is still limited. In this work we analyze P. fermentans EV production during growth in two different media containing urea (YCU) or methionine (YCM) where yeast-like or pseudohyphal morphology are produced. We developed a procedure to extract EV from the neighboring biofilm which is faster and more efficient as compared to the widely used ultracentrifugation method. Differences in morphology and RNA content of EV suggest that they might have an active role during dimorphic transition as response to the growth conditions. Our findings are coherent with a general state of hypoxic stress of the pseudohyphal cells.

Keywords: Pichia fermentans; cell communication; dimorphic transition; extracellular vesicles.

MeSH terms

  • Biofilms*
  • Culture Media
  • Extracellular Vesicles / metabolism*
  • Extracellular Vesicles / ultrastructure
  • Fungal Proteins / metabolism
  • MicroRNAs / metabolism
  • Microbial Viability
  • Pichia / cytology
  • Pichia / physiology*
  • Pichia / ultrastructure
  • RNA, Fungal / metabolism

Substances

  • Culture Media
  • Fungal Proteins
  • MicroRNAs
  • RNA, Fungal