title: Dynamics of cullin-RING ubiquitin ligase network revealed by systematic quantitative proteomics.
abstract: Dynamic reorganization of signaling systems frequently accompanies pathway perturbations, yet quantitative studies of network remodeling by pathway stimuli are lacking. Here, we report the development of a quantitative proteomics platform centered on multiplex absolute quantification (AQUA) technology to elucidate the architecture of the cullin-RING ubiquitin ligase (CRL) network and to evaluate current models of dynamic CRL remodeling. Current models suggest that CRL complexes are controlled by cycles of CRL deneddylation and CAND1 binding. Contrary to expectations, acute CRL inhibition with MLN4924, an inhibitor of the NEDD8-activating enzyme, does not result in a global reorganization of the CRL network. Examination of CRL complex stoichiometry reveals that, independent of cullin neddylation, a large fraction of cullins are assembled with adaptor modules, whereas only a small fraction are associated with CAND1. These studies suggest an alternative model of CRL dynamicity where the abundance of adaptor modules, rather than cycles of neddylation and CAND1 binding, drives CRL network organization.
authors.affiliation: Department of Pathology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Bennett
authors.forename: John
authors.initials: J
authors.lastname: Rush
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
authors.forename: J Wade
authors.initials: JW
authors.lastname: Harper
mesh_terms: D002460:Cell Line; D044842:Cullin Proteins / Q000737:chemistry / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology; D000075687:NEDD8 Protein; D011499:Protein Processing, Post-Translational; D040901:Proteomics; D011743:Pyrimidines / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism; D014452:Ubiquitins / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: C486317:CAND1 protein, human; C100318:Cullin 1; D044842:Cullin Proteins; D003517:Cyclopentanes; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D011743:Pyrimidines; D014157:Transcription Factors; D014452:Ubiquitins; C539933:pevonedistat