PXD030849

PXD030849 is an original dataset announced via ProteomeXchange.

Title	Cullin-independent recognition of HHARI substrates by a dynamic RBR catalytic domain
Description	RING-Between-RING (RBR) E3 ligases mediate ubiquitin transfer through an obligate E3- ubiquitin thioester intermediately prior to substrate ubiquitination. While RBRs share a conserved catalytic module, substrate recruitment mechanisms remain enigmatic and the relevant domains have yet to be identified for any member of the class. Here we characterize the interaction between the auto-inhibited RBR, HHARI (AriH1), and its target protein, 4EHP, using a combination of XL-MS, HDX-MS, NMR, and biochemical studies. The results show that 1) a di-aromatic surface on the catalytic HHARI Rcat domain forms a binding platform for substrates and 2) a phospho-mimetic mutation on the auto-inhibitory Ariadne domain of HHARI promotes release and reorientation of Rcat for transthiolation and substrate modification. The findings represent the first identification of a direct binding interaction between a RING-Between-RING ligase and its substrate.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:48:16.550.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Alex Zelter
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-09 11:45:29	ID requested
1	2023-05-24 10:58:25	announced
⏵ 2	2023-11-14 08:48:17	announced	2023-11-14: Updated project metadata.

submitter keyword: Kojak,Cross-linking Mass Spectrometry, ProXL, Skyline, Cullin, XL-MS, HHARI, ubiquitin, RBR, E3 ligase

Rachel E. Klevit
contact affiliation	Department of Biochemistry, University of Washington, Seattle, WA, USA
contact email	klevit@uw.edu
lab head
Alex Zelter
contact affiliation	Department of Biochemistry, University of Washington, Seattle
contact email	azelter@uw.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD030849
     1. Label: PRIDE project
     2. Name: Cullin-independent recognition of HHARI substrates by a dynamic RBR catalytic domain