Using Caenorhabditis elegans to screen for tissue-specific chaperone interactions

Shiran Dror, Tomer D. Meidan, Ido Karady, Anat Ben-Zvi

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Correct folding and assembly of proteins and protein complexes are essential for cellular function. Cells employ quality control pathways that correct, sequester or eliminate damaged proteins to maintain a healthy proteome, thus ensuring cellular proteostasis and preventing further protein damage. Because of redundant functions within the proteostasis network, screening for detectable phenotypes using knockdown or mutations in chaperone-encoding genes in the multicellular organism Caenorhabditis elegans results in the detection of minor or no phenotypes in most cases. We have developed a targeted screening strategy to identify chaperones required for a specific function and thus bridge the gap between phenotype and function. Specifically, we monitor novel chaperone interactions using RNAi synthetic interaction screens, knocking-down chaperone expression, one chaperone at a time, in animals carrying a mutation in a chaperone-encoding gene or over-expressing a chaperone of interest. By disrupting two chaperones that individually present no gross phenotype, we can identify chaperones that aggravate or expose a specific phenotype when both perturbed. We demonstrate that this approach can identify specific sets of chaperones that function together to modulate the folding of a protein or protein complexes associated with a given phenotype.

Original languageEnglish
Article numbere61140
Pages (from-to)1-12
Number of pages12
JournalJournal of Visualized Experiments
Volume2020
Issue number160
DOIs
StatePublished - 1 Jan 2020

Keywords

  • Biology
  • Caenorhabditis elegans
  • Chaperone
  • Genetic interactions
  • Issue 160
  • Proteostasis
  • RNAi
  • Screen
  • Temperature-sensitive

ASJC Scopus subject areas

  • Neuroscience (all)
  • Chemical Engineering (all)
  • Biochemistry, Genetics and Molecular Biology (all)
  • Immunology and Microbiology (all)

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