Faculty Collaboration Database

UBCH7 reactivity profile reveals parkin and HHARI to be RING/HECT hybrids. Nature 2011 Jun 02;474(7349):105-8

Date

05/03/2011

Pubmed ID

21532592

Pubmed Central ID

PMC3444301

DOI

10.1038/nature09966

Scopus ID

2-s2.0-79957949190 (requires institutional sign-in at Scopus site)   418 Citations

Abstract

Although the functional interaction between ubiquitin-conjugating enzymes (E2s) and ubiquitin ligases (E3s) is essential in ubiquitin (Ub) signalling, the criteria that define an active E2-E3 pair are not well established. The human E2 UBCH7 (also known as UBE2L3) shows broad specificity for HECT-type E3s, but often fails to function with RING E3s in vitro despite forming specific complexes. Structural comparisons of inactive UBCH7-RING complexes with active UBCH5-RING complexes reveal no defining differences, highlighting a gap in our understanding of Ub transfer. Here we show that, unlike many E2s that transfer Ub with RINGs, UBCH7 lacks intrinsic, E3-independent reactivity with lysine, explaining its preference for HECTs. Despite lacking lysine reactivity, UBCH7 exhibits activity with the RING-in-between-RING (RBR) family of E3s that includes parkin (also known as PARK2) and human homologue of ariadne (HHARI; also known as ARIH1). Found in all eukaryotes, RBRs regulate processes such as translation and immune signalling. RBRs contain a canonical C3HC4-type RING, followed by two conserved Cys/His-rich Zn(2+)-binding domains, in-between-RING (IBR) and RING2 domains, which together define this E3 family. We show that RBRs function like RING/HECT hybrids: they bind E2s via a RING domain, but transfer Ub through an obligate thioester-linked Ub (denoted ∼Ub), requiring a conserved cysteine residue in RING2. Our results define the functional cadre of E3s for UBCH7, an E2 involved in cell proliferation and immune function, and indicate a novel mechanism for an entire class of E3s.

Author List

Wenzel DM, Lissounov A, Brzovic PS, Klevit RE

Author

Dawn M. Wenzel PhD Assistant Professor in the Biochemistry department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Amino Acid Sequence
Carrier Proteins
Catalytic Domain
Cysteine
Humans
Lysine
Molecular Sequence Data
Mutant Chimeric Proteins
Protein Binding
Protein Structure, Tertiary
Sequence Alignment
Ubiquitin
Ubiquitin-Conjugating Enzymes
Ubiquitin-Protein Ligases