ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombe.
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.
Matsuyama
Akihisa
A
Chemical Genetics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan.
Arai
Ritsuko
R
Yashiroda
Yoko
Y
Shirai
Atsuko
A
Kamata
Ayako
A
Sekido
Shigeko
S
Kobayashi
Yumiko
Y
Hashimoto
Atsushi
A
Hamamoto
Makiko
M
Hiraoka
Yasushi
Y
Horinouchi
Sueharu
S
Yoshida
Minoru
M
D000935:Antifungal Agents / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D015723:Gene Library; D023061:Gene Order / Q000235:genetics*; D005800:Genes, Fungal; D020407:Internet; D028884:Karyopherins / Q000037:antagonists & inhibitors / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics*; D040901:Proteomics / Q000379:methods; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics*; D000097606:Exportin 1 Protein
D000935:Antifungal Agents; D005231:Fatty Acids, Unsaturated; D028884:Karyopherins; D008164:Luminescent Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D000097606:Exportin 1 Protein; C038753:leptomycin B