Image analysis reveals that Escherichia coli RecA protein consists of two domains.
The Escherichia coli RecA protein catalyzes homologous genetic recombination by forming helical polymers around DNA molecules. These polymers have an ATPase activity, which is essential for the movement of strands between two DNA molecules. One obstacle to structural studies of the RecA filament has been that the ATPase results in a dynamical polymer containing a mixture of states with respect to the bound ATP and its hydrolytic products. We have formed filaments which are trapped in the ADP-Pi state by substituting AIF4- for the Pi, and have used these stable filaments to generate a three-dimensional reconstruction from electron micrographs. The resolution of the reconstruction is sufficient to resolve the 38-k RecA subunit into two nearly equal domains. This reconstruction provides the most detailed view yet of the RecA protein, and serves as a framework within which existing biochemical data on RecA can be understood.
Yu
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Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511.
Egelman
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EH
D010584:Bacteriophage phi X 174 / Q000378:metabolism; D003196:Computer Graphics; D004279:DNA, Viral / Q000378:metabolism / Q000648:ultrastructure; D004926:Escherichia coli / Q000378:metabolism; D005583:Fourier Analysis; D046911:Macromolecular Substances; D008854:Microscopy, Electron / Q000379:methods; D008958:Models, Molecular; D011487:Protein Conformation; D011938:Rec A Recombinases / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure*
D004279:DNA, Viral; D046911:Macromolecular Substances; D011938:Rec A Recombinases