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TitleStructural basis for DNA targeting by the Tn7 transposon.
Journal, issue, pagesNat Struct Mol Biol, Vol. 29, Issue 2, Page 143-151, Year 2022
Publish dateFeb 16, 2022
AuthorsYao Shen / Josue Gomez-Blanco / Michael T Petassi / Joseph E Peters / Joaquin Ortega / Alba Guarné /
PubMed AbstractTn7 transposable elements are unique for their highly specific, and sometimes programmable, target-site selection mechanisms and precise insertions. All the elements in the Tn7 family utilize an AAA+ ...Tn7 transposable elements are unique for their highly specific, and sometimes programmable, target-site selection mechanisms and precise insertions. All the elements in the Tn7 family utilize an AAA+ adaptor (TnsC) to coordinate target-site selection with transpososome assembly and to prevent insertions at sites already containing a Tn7 element. Owing to its multiple functions, TnsC is considered the linchpin in the Tn7 element. Here we present the high-resolution cryo-EM structure of TnsC bound to DNA using a gain-of-function variant of the protein and a DNA substrate that together recapitulate the recruitment to a specific DNA target site. TnsC forms an asymmetric ring on target DNA that segregates target-site selection and interaction with the paired-end complex to opposite faces of the ring. Unlike most AAA+ ATPases, TnsC uses a DNA distortion to find the target site but does not remodel DNA to activate transposition. By recognizing pre-distorted substrates, TnsC creates a built-in regulatory mechanism where ATP hydrolysis abolishes ring formation proximal to an existing element. This work unveils how Tn7 and Tn7-like elements determine the strict spacing between the target and integration sites.
External linksNat Struct Mol Biol / PubMed:35173349
MethodsEM (single particle) / X-ray diffraction
Resolution3.2 - 3.56 Å
Structure data

23757

7mcs

EMDB-23757, PDB-7mcs:
Cryo-electron microscopy structure of TnsC(1-503)A225V bound to DNA
Method: EM (single particle) / Resolution: 3.56 Å

PDB-7mbw:
Crystal structure of TnsC(1-503)A225V
Method: X-RAY DIFFRACTION / Resolution: 3.2 Å

Chemicals

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

ChemComp-MG:
Unknown entry

ChemComp-HOH:
WATER / Water

ChemComp-ANP:
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMP-PNP, energy-carrying molecule analogue*YM

Source
  • escherichia coli (E. coli)
  • synthetic construct (others)
KeywordsDNA BINDING PROTEIN / AAA+ ATPase / Transposition / DNA binding / DNA BINDING PROTEIN/DNA / Complex / DNA BINDING PROTEIN-DNA complex

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