Skp1 forms multiple protein complexes, including RAVE, a regulator of V-ATPase assembly
- 9 March 2001
- journal article
- Published by Springer Nature in Nature Cell Biology
- Vol. 3 (4) , 384-391
- https://doi.org/10.1038/35070067
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.Keywords
This publication has 34 references indexed in Scilit:
- ROC1, a Homolog of APC11, Represents a Family of Cullin Partners with an Associated Ubiquitin Ligase ActivityPublished by Elsevier ,2014
- SCF and Cullin/RING H2-Based Ubiquitin LigasesAnnual Review of Cell and Developmental Biology, 1999
- Cdc53/cullin and the essential Hrt1 RING-H2 subunit of SCF define a ubiquitin ligase module that activates the E2 enzyme Cdc34Genes & Development, 1999
- Rbx1, a Component of the VHL Tumor Suppressor Complex and SCF Ubiquitin LigaseScience, 1999
- Recruitment of a ROC1–CUL1 Ubiquitin Ligase by Skp1 and HOS to Catalyze the Ubiquitination of IκBαMolecular Cell, 1999
- Combinatorial control in ubiquitin-dependent proteolysis: don't Skp the F-box hypothesisTrends in Genetics, 1998
- Cdc53 is a scaffold protein for multiple Cdc34/Skp1/F-box protein complexes that regulate cell division and methionine biosynthesis in yeastGenes & Development, 1998
- F-Box Proteins Are Receptors that Recruit Phosphorylated Substrates to the SCF Ubiquitin-Ligase ComplexCell, 1997
- Toward a functional analysis of the yeast genome through exhaustive two-hybrid screensNature Genetics, 1997
- A protein linkage map of Escherichia coli bacteriophage T7Nature Genetics, 1996