Ras hyperactivation versus overexpression: Lessons from Ras dynamics in Candida albicans

Vavilala A Pratyusha, Guiliana Soraya Victoria, Mohammad Firoz Khan, Dominic T Haokip, Bhawna Yadav, Nibedita Pal, Subhash Chandra Sethi, Priyanka Jain, Sneh Lata Singh, Sobhan Sen, Sneha Sudha Komath

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Abstract

Ras signaling in response to environmental cues is critical for cellular morphogenesis in eukaryotes. This signaling is tightly regulated and its activation involves multiple players. Sometimes Ras signaling may be hyperactivated. In C. albicans, a human pathogenic fungus, we demonstrate that dynamics of hyperactivated Ras1 (Ras1G13V or Ras1 in Hsp90 deficient strains) can be reliably differentiated from that of normal Ras1 at (near) single molecule level using fluorescence correlation spectroscopy (FCS). Ras1 hyperactivation results in significantly slower dynamics due to actin polymerization. Activating actin polymerization by jasplakinolide can produce hyperactivated Ras1 dynamics. In a sterol-deficient hyperfilamentous GPI mutant of C. albicans too, Ras1 hyperactivation results from Hsp90 downregulation and causes actin polymerization. Hyperactivated Ras1 co-localizes with G-actin at the plasma membrane rather than with F-actin. Depolymerizing actin with cytochalasin D results in faster Ras1 dynamics in these and other strains that show Ras1 hyperactivation. Further, ergosterol does not influence Ras1 dynamics.

Original languageEnglish
Article number5248
JournalScientific Reports
Volume8
Early online date27 Mar 2018
DOIs
Publication statusPublished - 2018

Keywords

  • Journal Article
  • Cell signalling
  • Fluorescence specrometry
  • Fungal biology
  • Membrane biophysics

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    Pratyusha, V. A., Victoria, G. S., Khan, M. F., Haokip, D. T., Yadav, B., Pal, N., Sethi, S. C., Jain, P., Singh, S. L., Sen, S., & Komath, S. S. (2018). Ras hyperactivation versus overexpression: Lessons from Ras dynamics in Candida albicans. Scientific Reports, 8, [5248]. https://doi.org/10.1038/s41598-018-23187-8