Functional 3D architecture in an intrinsically disordered E3 ligase domain facilitates ubiquitin transfer

TY - JOUR

T1 - Functional 3D architecture in an intrinsically disordered E3 ligase domain facilitates ubiquitin transfer

AU - Murphy, Paul

AU - Xu, Yingqi

AU - Rouse, Sarah L.

AU - Jaffray, Ellis G.

AU - Plechanovova, Anna

AU - Matthews, Steve J.

AU - Penedo, J. Carlos

AU - Hay, Ronald

N1 - This work was supported by the following grants: Wellcome Trust Investigator Award (098391/Z/12/Z) to R.T.H., Wellcome Trust Studentship (109113/Z/15/Z) to P.M., Wellcome Trust Collaborative Award (215539) and multiuser equipment grant (104833) to S.J.M. Additionally J.C.P. thanks the Scottish Universities Physics Alliance (SUPA) and the University of St. Andrews for financial support. We would also like to thank Michael Tatham for his contribution to data analysis and Federico Pelisch for his feedback on the manuscript. In addition, we would like to acknowledge Professor Takeshi Urano (Shimane University School of Medicine, Japan) for kindly providing the monoclonal antibody to RNF4.

PY - 2020/7/30

Y1 - 2020/7/30

N2 - The human genome contains an estimated 600 ubiquitin E3 ligases, many of which are single-subunit E3s (ssE3s) that can bind to both substrate and ubiquitin-loaded E2 (E2~Ub). Within ssE3s structural disorder tends to be located in substrate binding and domain linking regions. RNF4 is a ssE3 ligase with a C-terminal RING domain and disordered N-terminal region containing SUMO Interactions Motifs (SIMs) required to bind SUMO modified substrates. Here we show that, although the N-terminal region of RNF4 bears no secondary structure, it maintains a compact global architecture primed for SUMO interaction. Segregated charged regions within the RNF4 N-terminus promote compaction, juxtaposing RING domain and SIMs to facilitate substrate ubiquitination. Mutations that induce a more extended shape reduce ubiquitination activity. Our result offer insight into a key step in substrate ubiquitination by a member of the largest ubiquitin ligase subtype and reveal how a defined architecture within a disordered region contributes to E3 ligase function.

AB - The human genome contains an estimated 600 ubiquitin E3 ligases, many of which are single-subunit E3s (ssE3s) that can bind to both substrate and ubiquitin-loaded E2 (E2~Ub). Within ssE3s structural disorder tends to be located in substrate binding and domain linking regions. RNF4 is a ssE3 ligase with a C-terminal RING domain and disordered N-terminal region containing SUMO Interactions Motifs (SIMs) required to bind SUMO modified substrates. Here we show that, although the N-terminal region of RNF4 bears no secondary structure, it maintains a compact global architecture primed for SUMO interaction. Segregated charged regions within the RNF4 N-terminus promote compaction, juxtaposing RING domain and SIMs to facilitate substrate ubiquitination. Mutations that induce a more extended shape reduce ubiquitination activity. Our result offer insight into a key step in substrate ubiquitination by a member of the largest ubiquitin ligase subtype and reveal how a defined architecture within a disordered region contributes to E3 ligase function.

KW - Enzyme mechanisms

KW - Molecular conformation

KW - Solution-state NMR

KW - Ubiquitylation

UR - https://www.scopus.com/pages/publications/85088807112

U2 - 10.1038/s41467-020-17647-x

DO - 10.1038/s41467-020-17647-x

M3 - Article

C2 - 32733036

SN - 2041-1723

VL - 11

JO - Nature Communications

JF - Nature Communications

M1 - 3807

ER -