Nuclear export signal consensus sequences defined using a localization-based yeast selection system

Shunichi Kosugi, Masako Hasebe, Masaru Tomita, Hiroshi Yanagawa

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Proteins bearing nuclear export signals (NESs) are translocated to the cytoplasm from the nucleus mainly through the CRM1-dependent pathway. However, the NES consensus sequence remains poorly defined, and there are currently no high-throughput methods for identifying NESs. In this study, we report the development of an efficient yeast selection system for detecting nuclear export activity as well as several reliable NES consensus sequences identified using this method. Our selection system is based on the nuclear export-dependent rescue of Tys1p, an essential cytoplasmic protein that has been artificially localized to the nucleus in a haploid Δtys1 knockout strain. A screen of a random peptide library revealed 101 distinct CRM1-dependent NESs, which were classified into six patterns according to the conserved hydrophobic spacing. By combining mutational analyses, we have defined new NES consensus sequences with more specific and redundant residues than the traditional consensus sequence, which are consistent with most experimentally confirmed NESs. These NES consensus sequences should help identify functional NESs, and our selection system can be used to identify other targeting signals or proteins imported to specific subcellular compartments.

Original languageEnglish
Pages (from-to)2053-2062
Number of pages10
JournalTraffic
Volume9
Issue number12
DOIs
Publication statusPublished - 2008 Nov 20

Keywords

  • CRM1
  • Consensus
  • Nuclear export signal
  • Transportin

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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