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Bacteriophage φ29 scaffolding protein gp7 before and after prohead assembly

Nature Structural & Molecular Biology volume 10, pages 572–576 (2003)Cite this article

Abstract

Three-dimensional structures of the double-stranded DNA bacteriophage φ29 scaffolding protein (gp7) before and after prohead assembly have been determined at resolutions of 2.2 and 2.8 Å, respectively. Both structures are dimers that resemble arrows, with a four-helix bundle composing the arrowhead and a coiled coil forming the tail. The structural resemblance of gp7 to the yeast transcription factor GCN4 suggests a DNA-binding function that was confirmed by native gel electrophoresis. DNA binding to gp7 may have a role in mediating the structural transition from prohead to mature virus and scaffold release. A cryo-EM analysis indicates that gp7 is arranged inside the capsid as a series of concentric shells. The position of the higher density features in these shells correlates with the positions of hexamers in the equatorial region of the capsid, suggesting that gp7 may regulate formation of the prolate head through interactions with these hexamers.

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Figure 1: Assembly pathway of bacteriophage φ29.
Figure 2: Stereo diagrams of the gp7 structure.
Figure 3: Cryo-EM density of the prohead scaffold lattice.
Figure 4: Native agarose gel stained with ethidium bromide.

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Acknowledgements

We thank P. Jardine for helpful discussions and encouragement, and S. Wilder and C. Towell for help in preparation of the manuscript. The work was supported by a US National Science Foundation grant to M.G.R., National Institutes of Health grants to D.L.A. and a National Institutes of Health postdoctoral fellowship to M.C.M. We are also grateful for a grant from the Keck Foundation that provided for the purchase of a Philips CM300 FEG electron microscope, as well as for a Purdue University reinvestment grant to support structural biology.

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Authors and Affiliations

  1. Department of Biological Sciences, Purdue University, 915 West State Street, West Lafayette, 47907-2054, Indiana, USA

    Marc C Morais, Shuji Kanamaru & Michael G Rossmann

  2. Department of Oral Science, 18-242 Moos Tower, University of Minnesota, Minneapolis, 55455, Minnesota, USA

    Mohammed O Badasso & Jaya S Koti

  3. Department of Molecular Pharmacology & Experimental Therapeutics, Mayo Clinic and Foundation, Rochester, 55905, Minnesota, USA

    Barbara A L Owen & Cynthia T McMurray

  4. Departments of Microbiology and Oral Science, 18-242 Moos Tower, University of Minnesota, Minneapolis, 55455, Minnesota, USA

    Dwight L Anderson

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Correspondence to Michael G Rossmann.

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Morais, M., Kanamaru, S., Badasso, M. et al. Bacteriophage φ29 scaffolding protein gp7 before and after prohead assembly. Nat Struct Mol Biol 10, 572–576 (2003). https://doi.org/10.1038/nsb939

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