Structural mechanisms of autoinhibition and substrate recognition by the ubiquitin ligase HACE1
2024 | journal article. A publication with affiliation to the University of Göttingen.
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Structural mechanisms of autoinhibition and substrate recognition by the ubiquitin ligase HACE1
Düring, J.; Wolter, M.; Toplak, J. J.; Torres, C.; Dybkov, O.; Fokkens, T. J. & Bohnsack, K. E. et al. (2024)
Nature Structural & Molecular Biology, 31(2) pp. 364-377. DOI: https://doi.org/10.1038/s41594-023-01203-4
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Details
- Authors
- Düring, Jonas; Wolter, Madita; Toplak, Julia J.; Torres, Camilo; Dybkov, Olexandr; Fokkens, Thornton J.; Bohnsack, Katherine E.; Urlaub, Henning; Steinchen, Wieland; Dienemann, Christian; Lorenz, Sonja
- Abstract
- Ubiquitin ligases (E3s) are pivotal specificity determinants in the ubiquitin system by selecting substrates and decorating them with distinct ubiquitin signals. However, structure determination of the underlying, specific E3-substrate complexes has proven challenging owing to their transient nature. In particular, it is incompletely understood how members of the catalytic cysteine-driven class of HECT-type ligases (HECTs) position substrate proteins for modification. Here, we report a cryogenic electron microscopy (cryo-EM) structure of the full-length human HECT HACE1, along with solution-based conformational analyses by small-angle X-ray scattering and hydrogen–deuterium exchange mass spectrometry. Structure-based functional analyses in vitro and in cells reveal that the activity of HACE1 is stringently regulated by dimerization-induced autoinhibition. The inhibition occurs at the first step of the catalytic cycle and is thus substrate-independent. We use mechanism-based chemical crosslinking to reconstitute a complex of activated, monomeric HACE1 with its major substrate, RAC1, determine its structure by cryo-EM and validate the binding mode by solution-based analyses. Our findings explain how HACE1 achieves selectivity in ubiquitinating the active, GTP-loaded state of RAC1 and establish a framework for interpreting mutational alterations of the HACE1–RAC1 interplay in disease. More broadly, this work illuminates central unexplored aspects in the architecture, conformational dynamics, regulation and specificity of full-length HECTs.
- Issue Date
- 2024
- Journal
- Nature Structural & Molecular Biology
- Project
- EXC 2067: Multiscale Bioimaging
SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente
SFB 1565: Molekulare Mechanismen und Vernetzung von Prozessen der Genexpression
SFB 1565 | P12: Posttranslationale Regulation von Ribonukleasen und ihrer Funktionen in Prozessierung und Abbau unterschiedlicher Klassen von RNAs unter Zellstress - Working Group
- RG Urlaub (Bioanalytische Massenspektrometrie)
RG K. Bohnsack (RNA Metabolism)
RG Lorenz (Ubiquitin Signaling Specificity) - ISSN
- 1545-9993
- eISSN
- 1545-9985
- Language
- English