A gene signal amplifier platform for monitoring the unfolded protein response

Nat Chem Biol. 2020 May;16(5):520-528. doi: 10.1038/s41589-020-0497-x. Epub 2020 Mar 9.

Abstract

Gene expression in mammalian cells results from coordinated protein-driven processes guided by diverse mechanisms of regulation, including protein-protein interactions, protein localization, DNA modifications and chromatin rearrangement. Regulation of gene expression is particularly important in stress-response pathways. To address the need to monitor chromosomal gene expression generating a readily detectable signal output that recapitulates gene expression dynamics, we developed a gene signal amplifier platform that links transcriptional and post-translational regulation of a fluorescent output to the expression of a chromosomal target gene. We generated a multiplex reporter system for monitoring markers of the unfolded protein response, a complex signal transduction pathway that remodels gene expression in response to proteotoxic stress in the endoplasmic reticulum. By recapitulating the transcriptional and translational control mechanisms underlying the expression of a target gene with high sensitivity, this platform provides a technology for monitoring gene expression with superior sensitivity and dynamic resolution.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Activating Transcription Factor 6 / genetics
  • Chromosomes / genetics
  • Computational Biology / methods
  • Endoplasmic Reticulum
  • Endoribonucleases / genetics
  • Gene Expression Profiling / methods*
  • Genes, Reporter*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Protein Biosynthesis
  • Protein Serine-Threonine Kinases / genetics
  • Transcription, Genetic
  • Unfolded Protein Response / genetics*
  • eIF-2 Kinase / genetics

Substances

  • ATF6 protein, human
  • Activating Transcription Factor 6
  • Green Fluorescent Proteins
  • EIF2AK3 protein, human
  • ERN1 protein, human
  • Protein Serine-Threonine Kinases
  • eIF-2 Kinase
  • Endoribonucleases