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TitleCryo-EM reveals the transition of Arp2/3 complex from inactive to nucleation-competent state.
Journal, issue, pagesNat Struct Mol Biol, Vol. 27, Issue 11, Page 1009-1016, Year 2020
Publish dateAug 24, 2020
AuthorsMohammed Shaaban / Saikat Chowdhury / Brad J Nolen /
PubMed AbstractArp2/3 complex, a crucial actin filament nucleator, undergoes structural rearrangements during activation by nucleation-promoting factors (NPFs). However, the conformational pathway leading to the ...Arp2/3 complex, a crucial actin filament nucleator, undergoes structural rearrangements during activation by nucleation-promoting factors (NPFs). However, the conformational pathway leading to the nucleation-competent state is unclear due to lack of high-resolution structures of the activated state. Here we report a ~3.9 Å resolution cryo-EM structure of activated Schizosaccharomyces pombe Arp2/3 complex bound to the S. pombe NPF Dip1 and attached to the end of the nucleated actin filament. The structure reveals global and local conformational changes that allow the two actin-related proteins in Arp2/3 complex to mimic a filamentous actin dimer and template nucleation. Activation occurs through a clamp-twisting mechanism, in which Dip1 forces two core subunits in Arp2/3 complex to pivot around one another, shifting half of the complex into a new activated position. By showing how Dip1 stimulates activation, the structure reveals how NPFs can activate Arp2/3 complex in diverse cellular processes.
External linksNat Struct Mol Biol / PubMed:32839613 / PubMed Central
MethodsEM (single particle)
Resolution3.9 - 4.2 Å
Structure data

EMDB-21502, PDB-6w17:
Structure of Dip1-activated Arp2/3 complex with nucleated actin filament
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-21503, PDB-6w18:
Structure of S. pombe Arp2/3 complex in inactive state
Method: EM (single particle) / Resolution: 4.2 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

Source
  • schizosaccharomyces pombe (strain 972 / atcc 24843) (yeast)
  • Fission yeast (fission yeast)
  • Rabbit (rabbit)
  • death cap (death cap)
  • oryctolagus cuniculus (rabbit)
  • amanita phalloides (death cap)
KeywordsSTRUCTURAL PROTEIN / Arp2/3 / actin / Dip1 / cytoskeletal protein / actin regulator

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