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Structure paper

TitleStructures of a RAG-like transposase during cut-and-paste transposition.
Journal, issue, pagesNature, Vol. 575, Issue 7783, Page 540-544, Year 2019
Publish dateNov 13, 2019
AuthorsChang Liu / Yang Yang / David G Schatz /
PubMed AbstractTransposons have had a pivotal role in genome evolution and are believed to be the evolutionary progenitors of the RAG1-RAG2 recombinase, an essential component of the adaptive immune system in jawed ...Transposons have had a pivotal role in genome evolution and are believed to be the evolutionary progenitors of the RAG1-RAG2 recombinase, an essential component of the adaptive immune system in jawed vertebrates. Here we report one crystal structure and five cryo-electron microscopy structures of Transib, a RAG1-like transposase from Helicoverpa zea, that capture the entire transposition process from the apo enzyme to the terminal strand transfer complex with transposon ends covalently joined to target DNA, at resolutions of 3.0-4.6 Å. These structures reveal a butterfly-shaped complex that undergoes two cycles of marked conformational changes in which the 'wings' of the transposase unfurl to bind substrate DNA, close to execute cleavage, open to release the flanking DNA and close again to capture and attack target DNA. Transib possesses unique structural elements that compensate for the absence of a RAG2 partner, including a loop that interacts with the transposition target site and an accordion-like C-terminal tail that elongates and contracts to help to control the opening and closing of the enzyme and assembly of the active site. Our findings reveal the detailed reaction pathway of a eukaryotic cut-and-paste transposase and illuminate some of the earliest steps in the evolution of the RAG recombinase.
External linksNature / PubMed:31723264 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution3.01 - 4.6 Å
Structure data

EMDB-20452, PDB-6pqr:
Cryo-EM structure of HzTransib/intact TIR substrate DNA pre-reaction complex (PRC)
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-20453, PDB-6pqu:
Cryo-EM structure of HzTransib/nicked TIR substrate DNA pre-reaction complex (PRC)
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-20455, PDB-6pqx:
Cryo-EM structure of HzTransib/nicked TIR substrate DNA hairpin forming complex (HFC)
Method: EM (single particle) / Resolution: 4.6 Å

EMDB-20456, PDB-6pqy:
Cryo-EM structure of HzTransib/TIR DNA transposon end complex (TEC)
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-20457, PDB-6pr5:
Cryo-EM structure of HzTransib strand transfer complex (STC)
Method: EM (single particle) / Resolution: 3.3 Å

PDB-6pqn:
Crystal structure of HzTransib transposase
Method: X-RAY DIFFRACTION / Resolution: 3.01 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-PO4:
PHOSPHATE ION

ChemComp-GOL:
GLYCEROL

ChemComp-NA:
Unknown entry

ChemComp-HOH:
WATER

ChemComp-MG:
Unknown entry

ChemComp-K:
Unknown entry

ChemComp-CA:
Unknown entry

Source
  • helicoverpa zea (corn earworm)
KeywordsRECOMBINATION / RAG-like transposase / DDE family enzyme / Transib / Evolution of RAG / recombination/dna / Terminal inverted repeat. / recombination-dna complex / Strand transfer.

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