Flexibility of the rings: structural asymmetry in the DnaB hexameric helicase.
DnaB is the primary replicative helicase in Escherichia coli and the hexameric DnaB ring has previously been shown to exist in two states in the presence of nucleotides. In one, all subunits are equivalent, while in the other, there are two different subunit conformations resulting in a trimer of dimers. Under all conditions that we have used for electron microscopy, including the absence of nucleotide, some rings exist as trimers of dimers, showing that the symmetry of the DnaB hexamer can be broken prior to nucleotide binding. Three-dimensional reconstructions reveal that the N-terminal domain of DnaB makes two very different contacts with neighboring subunits in the trimer of dimers, but does not form a predicted dimer with a neighboring N-terminal domain. Within the trimer of dimers, the helicase domain exists in two alternate conformations, each of which can form symmetrical hexamers depending upon the nucleotide cofactor used. These results provide new information about the modular architecture and domain dynamics of helicases, and suggest, by comparison with the hexameric bacteriophage T7 gp4 and SV40 large T-antigen helicases, that a great structural and mechanistic diversity may exist among the hexameric helicases.
Department of Biochemistry and Molecular Genetics, University of Virginia Health Sciences Center, Jordan Hall, Box 800773, Charlottesville 22908-0733, USA.
Shixin
S
Yang
Xiong
X
Yu
Margaret S
MS
VanLoock
Maria J
MJ
Jezewska
Wlodzimierz
W
Bujalowski
Edward H
EH
Egelman
D001426:Bacterial Proteins; D004265:DNA Helicases / Q000737:chemistry* / Q000648:ultrastructure*; D019281:Dimerization; D053488:DnaB Helicases; D008958:Models, Molecular; D011487:Protein Conformation; D017434:Protein Structure, Tertiary
D001426:Bacterial Proteins; D004265:DNA Helicases; D053488:DnaB Helicases