Open AccessArchitecture of the bacteriophage T4 primosome: Electron microscopy studies of helicase (gp41) and primase (gp61)
Author(s) -
Mona Trempe Norcum,
Janet A. Warrington,
Michelle M. Spiering,
Faoud T. Ishmael,
Michael A. Trakselis,
Stephen J. Benkovic
Publication year - 2005
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0500713102
Subject(s) - primase , random hexamer , trimer , helicase , dna , bacteriophage , electron microscope , biology , biophysics , replisome , crystallography , dna replication , chemistry , biochemistry , dimer , physics , gene , reverse transcriptase , rna , organic chemistry , escherichia coli , circular bacterial chromosome , optics
Replication of DNA requires helicase and primase activities as part of a primosome assembly. In bacteriophage T4, helicase and primase are separate polypeptides for which little structural information is available and whose mechanism of association within the primosome is not yet understood. Three-dimensional structural information is provided here by means of reconstructions from electron microscopic images. Structures have been calculated for complexes of each of these proteins with ssDNA in the presence of MgATPgammaS. Both the helicase (gp41) and primase (gp61) complexes are asymmetric hexagonal rings. The gp41 structure suggests two distinct forms that have been termed "open" and "closed." The gp61 structure is clearly a six-membered ring, which may be a trimer of dimers or a traditional hexamer of monomers. This structure provides conclusive evidence for an oligomeric primase-to-ssDNA stoichiometry of 6:1.