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TitleElucidating the Architectural dynamics of MuB filaments in bacteriophage Mu DNA transposition.
Journal, issue, pagesNat Commun, Vol. 15, Issue 1, Page 6445, Year 2024
Publish dateJul 31, 2024
AuthorsXiaolong Zhao / Yongxiang Gao / Qingguo Gong / Kaiming Zhang / Shanshan Li /
PubMed AbstractMuB is a non-specific DNA-binding protein and AAA+ ATPase that significantly influences the DNA transposition process of bacteriophage Mu, especially in target DNA selection for transposition. While ...MuB is a non-specific DNA-binding protein and AAA+ ATPase that significantly influences the DNA transposition process of bacteriophage Mu, especially in target DNA selection for transposition. While studies have established the ATP-dependent formation of MuB filament as pivotal to this process, the high-resolution structure of a full-length MuB protomer and the underlying molecular mechanisms governing its oligomerization remain elusive. Here, we use cryo-EM to obtain a 3.4-Å resolution structure of the ATP(+)-DNA(+)-MuB helical filament, which encapsulates the DNA substrate within its axial channel. The structure categorizes MuB within the initiator clade of the AAA+ protein family and precisely locates the ATP and DNA binding sites. Further investigation into the oligomeric states of MuB show the existence of various forms of the filament. These findings lead to a mechanistic model where MuB forms opposite helical filaments along the DNA, exposing potential target sites on the bare DNA and then recruiting MuA, which stimulates MuB's ATPase activity and disrupts the previously formed helical structure. When this happens, MuB generates larger ring structures and dissociates from the DNA.
External linksNat Commun / PubMed:39085263 / PubMed Central
MethodsEM (helical sym.) / EM (single particle)
Resolution3.4 - 7.72 Å
Structure data

38695
EMDB entry, No image

8xvb

EMDB-38695, PDB-8xvb:
Cryo-EM structure of ATP-DNA-MuB filaments
Method: EM (helical sym.) / Resolution: 3.4 Å

38696
EMDB entry, No image

8xvc

EMDB-38696, PDB-8xvc:
CryoEM structure of ADP-DNA-MuB conformation1
Method: EM (single particle) / Resolution: 4.32 Å

38697
EMDB entry, No image

8xvd

EMDB-38697, PDB-8xvd:
CryoEM structure of ADP-DNA-MuB conformation2
Method: EM (single particle) / Resolution: 4.43 Å


EMDB entry, No image

EMDB-38698: CryoEM structure of ADP-DNA-MuB conformation3
Method: EM (single particle) / Resolution: 6.91 Å


EMDB entry, No image

EMDB-38699: CryoEM structure of ADP-DNA-MuB conformation4
Method: EM (single particle) / Resolution: 7.72 Å

Chemicals

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

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

Source
  • escherichia phage mu (virus)
KeywordsVIRAL PROTEIN/DNA / VIRAL PROTEIN-DNA COMPLEX / VIRAL PROTEIN / MuB / MuB-ADP / Ring / ADP-MuB complex / D10 symmetry

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