Isolation and characterization of a novel F-box protein Pof10 in fission yeast.
The SCF complex is a type of ubiquitin-protein ligase (E3) that consists of invariable components, including Skp1, Cdc53/Cul1, and Rbx1, as well as variable components known as F-box proteins. Using a yeast two-hybrid system, we isolated six proteins that interact with Schizosaccharomyces pombe Skp1. Among them, Pof10 is a novel F-box protein consisting of 662 amino acids, harboring the F-box domain required for the binding to Skp1 and followed by four WD40 repeats. Overexpression of Pof10 in fission yeast resulted in loss of viability with marked morphological changes that are similar to those in pop1 mutant yeast. Coexpression of Skp1 with Pof10 prevented the lethality, suggesting that the lethality from Pof10 overexpression results from the sequestration of Skp1 from other F-box proteins including Pop1. Whereas most F-box proteins show rapid turnover, Pof10 has a remarkably long half-life in vivo and has been shown to be localized predominantly in cytoplasm. These results suggest that the stable F-box protein Pof10 might target abundant cytoplasmic proteins for degradation in fission yeast.
Ikebe
Chiho
C
Department of Molecular and Cellular Biology, Kyushu University, Fukuoka, Fukuoka, 812-8582, Japan.
Kominami
Kin-ichiro
K
Toda
Takashi
T
Nakayama
Kei-Ichi
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D018797:Cell Cycle Proteins / Q000378:metabolism; D044783:F-Box Proteins; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D017353:Gene Deletion; D006207:Half-Life; D008969:Molecular Sequence Data; D010453:Peptide Synthases / Q000378:metabolism; D044786:S-Phase Kinase-Associated Proteins; D044843:SKP Cullin F-Box Protein Ligases; D012568:Schizosaccharomyces / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development; D029702:Schizosaccharomyces pombe Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D013347:Subcellular Fractions / Q000378:metabolism
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D044783:F-Box Proteins; D005656:Fungal Proteins; C449205:Pof10 protein, S pombe; D044786:S-Phase Kinase-Associated Proteins; D029702:Schizosaccharomyces pombe Proteins; D044843:SKP Cullin F-Box Protein Ligases; D010453:Peptide Synthases