ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombe

Nat Biotechnol. 2006 Jul;24(7):841-7. doi: 10.1038/nbt1222. Epub 2006 Jun 25.

Abstract

Cloning of the entire set of an organism's protein-coding open reading frames (ORFs), or 'ORFeome', is a means of connecting the genome to downstream 'omics' applications. Here we report a proteome-scale study of the fission yeast Schizosaccharomyces pombe based on cloning of the ORFeome. Taking advantage of a recombination-based cloning system, we obtained 4,910 ORFs in a form that is readily usable in various analyses. First, we evaluated ORF prediction in the fission yeast genome project by expressing each ORF tagged at the 3' terminus. Next, we determined the localization of 4,431 proteins, corresponding to approximately 90% of the fission yeast proteome, by tagging each ORF with the yellow fluorescent protein. Furthermore, using leptomycin B, an inhibitor of the nuclear export protein Crm1, we identified 285 proteins whose localization is regulated by Crm1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antifungal Agents / pharmacology
  • Fatty Acids, Unsaturated / pharmacology
  • Gene Expression Regulation, Fungal / drug effects*
  • Gene Library*
  • Gene Order / genetics*
  • Genes, Fungal
  • Internet
  • Karyopherins / antagonists & inhibitors
  • Karyopherins / genetics
  • Luminescent Proteins / genetics
  • Open Reading Frames / genetics*
  • Proteomics / methods
  • Receptors, Cytoplasmic and Nuclear / antagonists & inhibitors
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces pombe Proteins / genetics*

Substances

  • Antifungal Agents
  • Fatty Acids, Unsaturated
  • Karyopherins
  • Luminescent Proteins
  • Receptors, Cytoplasmic and Nuclear
  • Saccharomyces cerevisiae Proteins
  • Schizosaccharomyces pombe Proteins
  • exportin 1 protein
  • leptomycin B