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TitleInsights into the ISG15 transfer cascade by the UBE1L activating enzyme.
Journal, issue, pagesNat Commun, Vol. 14, Issue 1, Page 7970, Year 2023
Publish dateDec 2, 2023
AuthorsIona Wallace / Kheewoong Baek / J Rajan Prabu / Ronnald Vollrath / Susanne von Gronau / Brenda A Schulman / Kirby N Swatek /
PubMed AbstractThe attachment of the ubiquitin-like protein ISG15 to substrates by specific E1-E2-E3 enzymes is a well-established signalling mechanism of the innate immune response. Here, we present a 3.45 Å ...The attachment of the ubiquitin-like protein ISG15 to substrates by specific E1-E2-E3 enzymes is a well-established signalling mechanism of the innate immune response. Here, we present a 3.45 Å cryo-EM structure of a chemically trapped UBE1L-UBE2L6 complex bound to activated ISG15. This structure reveals the details of the first steps of ISG15 recognition and UBE2L6 recruitment by UBE1L (also known as UBA7). Taking advantage of viral effector proteins from severe acute respiratory coronavirus 2 (SARS-CoV-2) and influenza B virus (IBV), we validate the structure and confirm the importance of the ISG15 C-terminal ubiquitin-like domain in the adenylation reaction. Moreover, biochemical characterization of the UBE1L-ISG15 and UBE1L-UBE2L6 interactions enables the design of ISG15 and UBE2L6 mutants with altered selectively for the ISG15 and ubiquitin conjugation pathways. Together, our study helps to define the molecular basis of these interactions and the specificity determinants that ensure the fidelity of ISG15 signalling during the antiviral response.
External linksNat Commun / PubMed:38042859 / PubMed Central
MethodsEM (single particle)
Resolution3.5 - 4.0 Å
Structure data

16891

8oif

EMDB-16891, PDB-8oif:
Structure of the UBE1L activating enzyme bound to ISG15 and UBE2L6
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-18589: Map of UBE1L activating enzyme bound to ISG15 (FL) and UBE2L6
Method: EM (single particle) / Resolution: 4.0 Å

Chemicals

ChemComp-AMP:
ADENOSINE MONOPHOSPHATE / AMP*YM / Adenosine monophosphate

Source
  • homo sapiens (human)
KeywordsANTIVIRAL PROTEIN / ubiquitin-like / ISG15 / interferon-stimulated genes / antiviral

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