Correlative single-molecule fluorescence barcoding of gene regulation in Saccharomyces cerevisiae

Methods. 2021 Sep:193:62-67. doi: 10.1016/j.ymeth.2020.10.009. Epub 2020 Oct 18.

Abstract

Most cells adapt to their environment by switching combinations of genes on and off through a complex interplay of transcription factor proteins (TFs). The mechanisms by which TFs respond to signals, move into the nucleus and find specific binding sites in target genes is still largely unknown. Single-molecule fluorescence microscopes, which can image single TFs in live cells, have begun to elucidate the problem. Here, we show that different environmental signals, in this case carbon sources, yield a unique single-molecule fluorescence pattern of foci of a key metabolic regulating transcription factor, Mig1, in the nucleus of the budding yeast, Saccharomyces cerevisiae. This pattern serves as a 'barcode' of the gene regulatory state of the cells which can be correlated with cell growth characteristics and other biological function.

Keywords: Energy sources; Growth; Single-molecule microscopy; Stress adaptation; Transcription factors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescence
  • Gene Expression Regulation
  • Gene Expression Regulation, Fungal
  • Repressor Proteins
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomyces cerevisiae* / genetics
  • Saccharomyces cerevisiae* / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • MIG1 protein, S cerevisiae
  • Repressor Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors