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TitlePrp19/Pso4 Is an Autoinhibited Ubiquitin Ligase Activated by Stepwise Assembly of Three Splicing Factors.
Journal, issue, pagesMol Cell, Vol. 69, Issue 6, Page 979-992.e6, Year 2018
Publish dateMar 15, 2018
AuthorsTales Rocha de Moura / Sina Mozaffari-Jovin / Csaba Zoltán Kibédi Szabó / Jana Schmitzová / Olexandr Dybkov / Constantin Cretu / Michael Kachala / Dmitri Svergun / Henning Urlaub / Reinhard Lührmann / Vladimir Pena /
PubMed AbstractHuman nineteen complex (NTC) acts as a multimeric E3 ubiquitin ligase in DNA repair and splicing. The transfer of ubiquitin is mediated by Prp19-a homotetrameric component of NTC whose elongated ...Human nineteen complex (NTC) acts as a multimeric E3 ubiquitin ligase in DNA repair and splicing. The transfer of ubiquitin is mediated by Prp19-a homotetrameric component of NTC whose elongated coiled coils serve as an assembly axis for two other proteins called SPF27 and CDC5L. We find that Prp19 is inactive on its own and have elucidated the structural basis of its autoinhibition by crystallography and mutational analysis. Formation of the NTC core by stepwise assembly of SPF27, CDC5L, and PLRG1 onto the Prp19 tetramer enables ubiquitin ligation. Protein-protein crosslinking of NTC, functional assays in vitro, and assessment of its role in DNA damage response provide mechanistic insight into the organization of the NTC core and the communication between PLRG1 and Prp19 that enables E3 activity. This reveals a unique mode of regulation for a complex E3 ligase and advances understanding of its dynamics in various cellular pathways.
External linksMol Cell / PubMed:29547724
MethodsSAS (X-ray synchrotron) / X-ray diffraction
Resolution1.605 - 2.8 Å
Structure data

SASDCT8:
Uncharacterized protein CTHT_0072540 (Core) from Chaetomium thermophilum
Method: SAXS/SANS

SASDCU8:
Uncharacterized protein CTHT_0072540 (WD40) from Chaetomium thermophilum
Method: SAXS/SANS

SASDCV8:
Uncharacterized protein CTHT_0072540 (Prp19 full-length) from Chaetomium thermophilum
Method: SAXS/SANS

PDB-5m88:
Spliceosome component
Method: X-RAY DIFFRACTION / Resolution: 2.8 Å

PDB-5m89:
Spliceosome component
Method: X-RAY DIFFRACTION / Resolution: 1.605 Å

PDB-5m8c:
Spliceosome component
Method: X-RAY DIFFRACTION / Resolution: 2.3 Å

Chemicals

ChemComp-HOH:
WATER

Source
  • chaetomium thermophilum (fungus)
  • chaetomium thermophilum (strain dsm 1495 / cbs 144.50 / imi 039719) (fungus)
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsSPLICING / Spliceosome

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