High-throughput identification of dominant negative polypeptides in yeast

Nat Methods. 2019 May;16(5):413-416. doi: 10.1038/s41592-019-0368-0. Epub 2019 Apr 8.

Abstract

Dominant negative polypeptides can inhibit protein function by binding to a wild-type subunit or by titrating a ligand. Here we use high-throughput sequencing of libraries composed of fragments of yeast genes to identify polypeptides that act in a dominant negative manner, in that they are depleted during cell growth. The method can uncover numerous inhibitory polypeptides for a protein and thereby define small inhibitory regions, even pinpointing individual residues with critical functional roles.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA-Binding Proteins / genetics
  • Gene Library
  • Genes, Dominant*
  • Genes, Fungal*
  • Heat-Shock Proteins / genetics
  • High-Throughput Nucleotide Sequencing / methods*
  • Peptides / genetics*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins / genetics
  • Sequence Analysis, DNA
  • Transcription Factors / genetics

Substances

  • DNA-Binding Proteins
  • HSF1 protein, S cerevisiae
  • Heat-Shock Proteins
  • Peptides
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • URA3 protein, S cerevisiae