title: Structure of HHARI, a RING-IBR-RING ubiquitin ligase: autoinhibition of an Ariadne-family E3 and insights into ligation mechanism.
abstract: A distinct mechanism for ubiquitin (Ub) ligation has recently been proposed for the RING1-IBR-RING2 (RBR) family of E3s: an N-terminal RING1 domain recruits a thioester-linked intermediate complex between Ub and the E2 UbcH7, and a structurally distinct C-terminal RING2 domain displays a catalytic cysteine required for Ub ligation. To obtain insights into RBR E3s, we determined the crystal structure of the human homolog of Ariadne (HHARI), which reveals the individual RING1, IBR, and RING2 domains embedded in superdomains involving sequences specific to the Ariadne RBR subfamily. The central IBR is flanked on one side by RING1, which is exposed and binds UbcH7. On the other side, a C-terminal autoinhibitory "Ariadne domain" masks the RING2 active site. Insights into RBR E3 mechanisms are provided by structure-based mutations that indicate distinct steps of relief from autoinhibition, Ub transfer from E2 to HHARI, and ligation from the HHARI cysteine to a terminal acceptor.
authors.affiliation: Howard Hughes Medical Institute, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Duda
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Olszewski
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Schuermann
authors.forename: Igor
authors.initials: I
authors.lastname: Kurinov
authors.forename: Darcie J
authors.initials: DJ
authors.lastname: Miller
authors.forename: Amanda
authors.initials: A
authors.lastname: Nourse
authors.forename: Arno F
authors.initials: AF
authors.lastname: Alpi
authors.forename: Brenda A
authors.initials: BA
authors.lastname: Schulman
mesh_terms: D002352:Carrier Proteins / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D044767:Ubiquitin-Protein Ligases / Q000037:antagonists & inhibitors / Q000737:chemistry*
chemical_list: D002352:Carrier Proteins; C419193:ARIH1 protein, human; D044767:Ubiquitin-Protein Ligases