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TitleStructural insights into Fe-S protein biogenesis by the CIA targeting complex.
Journal, issue, pagesNat Struct Mol Biol, Vol. 27, Issue 8, Page 735-742, Year 2020
Publish dateJul 6, 2020
AuthorsSusanne A Kassube / Nicolas H Thomä /
PubMed AbstractThe cytosolic iron-sulfur (Fe-S) assembly (CIA) pathway is required for the insertion of Fe-S clusters into cytosolic and nuclear client proteins, including many DNA replication and repair factors. ...The cytosolic iron-sulfur (Fe-S) assembly (CIA) pathway is required for the insertion of Fe-S clusters into cytosolic and nuclear client proteins, including many DNA replication and repair factors. The molecular mechanisms of client protein recognition and Fe-S cluster transfer remain unknown. Here, we report crystal structures of the CIA targeting complex (CTC), revealing that its CIAO2B subunit is centrally located and bridges CIAO1 and the client adaptor protein MMS19. Cryo-EM reconstructions of human CTC bound either to the DNA replication factor primase or to the DNA helicase DNA2, combined with biochemical, biophysical and yeast complementation assays, reveal an evolutionarily conserved, bipartite client recognition mode facilitated by CIAO1 and the structural flexibility of the MMS19 subunit. Unexpectedly, the primase Fe-S cluster is located ~70 Å away from the CTC reactive cysteine, implicating conformational dynamics of the CTC or additional maturation factors in the mechanism of Fe-S cluster transfer.
External linksNat Struct Mol Biol / PubMed:32632277
MethodsEM (single particle) / X-ray diffraction
Resolution2 - 15.8 Å
Structure data

EMDB-11016:
CTC-primase complex
Method: EM (single particle) / Resolution: 11.9 Å

EMDB-11017:
CTC-primase, body 1 (contains primase, CIAO1, CIAO2B, MMS19 CTD)
Method: EM (single particle) / Resolution: 8.8 Å

EMDB-11018:
CTC-primase, body 2 (contains MMS19 and CIAO2B)
Method: EM (single particle) / Resolution: 8.8 Å

EMDB-11019:
CTC-DNA2 complex
Method: EM (single particle) / Resolution: 15.8 Å

EMDB-11020:
CTC-DNA2, body 1 (contains DNA2, CIAO1, CIAO2B, and the CTD of MMS19)
Method: EM (single particle) / Resolution: 12.2 Å

EMDB-11021:
CTC-DNA2, body 2 (contains MMS19 and CIAO2B)
Method: EM (single particle) / Resolution: 12.2 Å

PDB-6tbl:
Crystal structure of MMS19(CTD)-CIAO1-CIAO2B CIA targeting complex
Method: X-RAY DIFFRACTION / Resolution: 2.65 Å

PDB-6tbn:
Crystal structure of CIAO1-CIAO2B CIA core complex
Method: X-RAY DIFFRACTION / Resolution: 2.0 Å

PDB-6tc0:
Crystal structure of MMS19-CIAO1-CIAO2B CIA targeting complex
Method: X-RAY DIFFRACTION / Resolution: 3.6 Å

Chemicals

ChemComp-K:
Unknown entry

ChemComp-EDO:
1,2-ETHANEDIOL

ChemComp-HOH:
WATER

ChemComp-NA:
Unknown entry

Source
  • Homo sapiens (human)
  • mus musculus (house mouse)
  • drosophila melanogaster (fruit fly)
KeywordsBIOSYNTHETIC PROTEIN / Fe-S cluster / Fe-S / CIA / CIA targeting complex / CIA core complex / CYTOSOLIC PROTEIN

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