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PMID:11283612
| Citation |
Seol, JH, Shevchenko, A, Shevchenko, A and Deshaies, RJ (2001) Skp1 forms multiple protein complexes, including RAVE, a regulator of V-ATPase assembly. Nat. Cell Biol. 3:384-91 |
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| Abstract |
SCF ubiquitin ligases are composed of Skp1, Cdc53, Hrt1 and one member of a large family of substrate receptors known as F-box proteins (FBPs). Here we report the identification, using sequential rounds of epitope tagging, affinity purification and mass spectrometry, of 16 Skp1 and Cdc53-associated proteins in budding yeast, including all components of SCF, 9 FBPs, Yjr033 (Rav1) and Ydr202 (Rav2). Rav1, Rav2 and Skp1 form a complex that we have named 'regulator of the (H+)-ATPase of the vacuolar and endosomal membranes' (RAVE), which associates with the V1 domain of the vacuolar membrane (H+)-ATPase (V-ATPase). V-ATPases are conserved throughout eukaryotes, and have been implicated in tumour metastasis and multidrug resistance, and here we show that RAVE promotes glucose-triggered assembly of the V-ATPase holoenzyme. Previous systematic genome-wide two-hybrid screens yielded 17 proteins that interact with Skp1 and Cdc53, only 3 of which overlap with those reported here. Thus, our results provide a distinct view of the interactions that link proteins into a comprehensive cellular network. |
| Links |
PubMed Online version:10.1038/35070067 |
| Keywords |
Cell Cycle Proteins/genetics; Cell Cycle Proteins/metabolism; Cullin Proteins; Cytoplasm/metabolism; Fungal Proteins/genetics; Fungal Proteins/metabolism; Glucose/metabolism; Holoenzymes/metabolism; Proton-Translocating ATPases/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; S-Phase Kinase-Associated Proteins; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins; Vacuolar Proton-Translocating ATPases |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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