Characterization of the cullin and F-box protein partner Skp1.
Skp1 interacts with cullins, F-box containing proteins, and forms a complex with cyclin A-Cdk2 in mammalian cells. Skp1 is also involved in diverse biological processes like degradation of key cell cycle regulators, glucose sensing, and kinetochore function. However, little is known about the structure and exact function of Skp1. Here we characterized the interaction between Skp1 and the F-box protein Skp2. We show that Skp1 can bind to Skp2 in vitro using recombinant proteins, and in vivo using the yeast two-hybrid system. Deletion analysis of Skp1 indicated that most of the Skp1 protein is required for binding to Skp2. In mammalian cell extracts, a large portion of Skp1 appears to associate with proteins other than Skp2. Biochemical analysis indicated that Skp1 is likely to be a flexible, non-spherical protein, and is capable of forming dimers.
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Department of Biochemistry, Hong Kong University of Science and Technology, Kowloon, China.
Arooz
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Yam
C H
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Chan
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IW
Lau
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Poon
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RY
D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D002850:Chromatography, Gel; D044842:Cullin Proteins; D019281:Dimerization; D015723:Gene Library; D006801:Humans; D007700:Kinetics; D008025:Ligases / Q000737:chemistry / Q000378:metabolism; D008322:Mammals; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D044786:S-Phase Kinase-Associated Proteins
D018797:Cell Cycle Proteins; C100318:Cullin 1; D044842:Cullin Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D044786:S-Phase Kinase-Associated Proteins; D008025:Ligases