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Structure paper

TitleAssembly of the Tn7 targeting complex by a regulated stepwise process.
Journal, issue, pagesMol Cell, Vol. 84, Issue 12, Page 2368-22381.e6, Year 2024
Publish dateJun 20, 2024
AuthorsYao Shen / Shreya S Krishnan / Michael T Petassi / Mark A Hancock / Joseph E Peters / Alba Guarné /
PubMed AbstractThe Tn7 family of transposons is notable for its highly regulated integration mechanisms, including programmable RNA-guided transposition. The targeting pathways rely on dedicated target selection ...The Tn7 family of transposons is notable for its highly regulated integration mechanisms, including programmable RNA-guided transposition. The targeting pathways rely on dedicated target selection proteins from the TniQ family and the AAA+ adaptor TnsC to recruit and activate the transposase at specific target sites. Here, we report the cryoelectron microscopy (cryo-EM) structures of TnsC bound to the TniQ domain of TnsD from prototypical Tn7 and unveil key regulatory steps stemming from unique behaviors of ATP- versus ADP-bound TnsC. We show that TnsD recruits ADP-bound dimers of TnsC and acts as an exchange factor to release one protomer with exchange to ATP. This loading process explains how TnsC assembles a heptameric ring unidirectionally from the target site. This unique loading process results in functionally distinct TnsC protomers within the ring, providing a checkpoint for target immunity and explaining how insertions at programmed sites precisely occur in a specific orientation across Tn7 elements.
External linksMol Cell / PubMed:38834067
MethodsEM (single particle)
Resolution3.32 - 3.88 Å
Structure data

40218

8glu

EMDB-40218, PDB-8glu:
CryoEM structure of TnsC(1-503) bound to TnsD(1-318) from E.coli Tn7
Method: EM (single particle) / Resolution: 3.57 Å

40221

8glw

EMDB-40221, PDB-8glw:
CryoEM structure of the TnsC(1-503)-TnsD(1-318)-DNA complex in a 7:2:1 stoichiometry from E. coli Tn7
Method: EM (single particle) / Resolution: 3.51 Å

40222

8glx

EMDB-40222, PDB-8glx:
CryoEM structure of the TnsC(1-503)-TnsD(1-318)-DNA complex in a 6:2:1 stoichiometry from E. coli Tn7
Method: EM (single particle) / Resolution: 3.88 Å

43138

8vcj

EMDB-43138, PDB-8vcj:
CryoEM structure of the TnsC(1-503)-TnsD(1-318)-DNA complex in a 7:2:1 stoichiometry from E. coli Tn7 bound to ATPgS and ADP
Method: EM (single particle) / Resolution: 3.32 Å

43140

8vct

EMDB-43140, PDB-8vct:
CyoEM structure of the TnsC(1-503)-TnsD(1-318)-DNA complex in a 6:2:1 stoichiometry from E. coli Tn7 bound to ATPgS and ADP
Method: EM (single particle) / Resolution: 3.83 Å

Chemicals

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

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

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

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

Source
  • escherichia coli (E. coli)
KeywordsDNA BINDING PROTEIN / Transposon / AAA+ ATPase / Oligomer / Complex / DNA BINDING PROTEIN/DNA / DNA BINDING PROTEIN-DNA complex / DNA-binding protein DNA complex

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