Polymorphic quaternary organization of the Bacillus subtilis bacteriophage SPP1 replicative helicase (G40 P).
The Bacillus subtilis bacteriophage SPP1 gene 40 product (G40P), which belongs to the DnaB-like family of helicases, is essential for SPP1 genome replication. The active form of the enzyme is the hexamer, capable of DNA unwinding with a 5' to 3' polarity fueled by the hydrolysis of a nucleoside 5'-triphosphate. We have used electron microscopy of negatively stained G40P samples and image processing techniques to study the structural characteristics of the hexameric assemblies of this protein. Our results provide the first low resolution data on a hexameric helicase of a Gram-positive bacterial origin. A novel approach has been adopted to analyze possible symmetry heterogeneities, an unsupervised method based on a neural network self-organizing algorithm, which has led to the detection of different subclasses of G40P views. Two different quaternary states of G40P homohexamers sharing a C3 symmetry organization have been found, as well as a minor class that seems to reflect an alternative C6 symmetry architecture. These forms show general features known for other hexameric helicases, such as the ring-like arrangement of monomers around a central hole. A clear structural handedness has also been detected in some of these forms. An analysis of these quaternary states and a model for the structural organization of G40P are presented.
Bárcena
M
M
Centro Nacional de Biotecnología-CSIC, Campus Universidad Autónoma, Madrid, 28049, Spain.
Martín
C S
CS
Weise
F
F
Ayora
S
S
Alonso
J C
JC
Carazo
J M
JM
D001412:Bacillus subtilis / Q000821:virology*; D001435:Bacteriophages / Q000201:enzymology*; D004265:DNA Helicases / Q000648:ultrastructure*; D004259:DNA-Directed DNA Polymerase / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron; D011487:Protein Conformation; D014764:Viral Proteins / Q000648:ultrastructure
D014764:Viral Proteins; C023140:DNA replicase; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases; C115901:G40P helicase protein, Bacteriophage SPP1