Discovery of pan autophagy inhibitors through a high-throughput screen highlights macroautophagy as an evolutionarily conserved process across 3 eukaryotic kingdoms

Research output: Contribution to journalArticlepeer-review


  • Piyush Mishra
  • Adrian N. Dauphinee
  • Arunika H. L. A. N. Gunawardena
  • Ravi Manjithaya

External organisations

  • Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research
  • Biology Department, Life Sciences Centre, Dalhousie University


Due to the involvement of macroautophagy/autophagy in different pathophysiological conditions such as infections, neurodegeneration and cancer, identification of novel small molecules that modulate the process is of current research and clinical interest. In this work, we developed a luciferase-based sensitive and robust kinetic high-throughput screen (HTS) of small molecules that modulate autophagic degradation of peroxisomes in the budding yeast Saccharomyces cerevisiae. Being a pathway-specific rather than a target-driven assay, we identified small molecule modulators that acted at key steps of autophagic flux. Two of the inhibitors, Bay11 and ZPCK, obtained from the screen were further characterized using secondary assays in yeast. Bay11 inhibited autophagy at a step before fusion with the vacuole whereas ZPCK inhibited the cargo degradation inside the vacuole. Furthermore, we demonstrated that these molecules altered the process of autophagy in mammalian cells as well. Strikingly, these molecules also modulated autophagic flux in a novel model plant, Aponogeton madagascariensis. Thus, using small molecule modulators identified by using a newly developed HTS autophagy assay, our results support that macroautophagy is a conserved process across fungal, animal and plant kingdoms.


Original languageEnglish
Pages (from-to)1556-1572
Number of pages17
Issue number9
Early online date9 Aug 2017
Publication statusE-pub ahead of print - 9 Aug 2017


  • Journal Article, autophagic flux, autophagy , high-throughput screening , kingdoms , peroxisome , programmed cell death , small molecules