Structure determination of the head-tail connector of bacteriophage φ29

A. A. Simpson, Petr Leiman, Y. Tao, Y. He, M. O. Badasso, P. J. Jardine, D. L. Anderson, M. G. Rossmann

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

The head-tail connector of bacteriophage φ29 is composed of 12 36 kDa subunits with 12-fold symmetry. It is the central component of a rotary motor that packages the genomic dsDNA into preformed proheads. This motor consists of the head-tail connector, surrounded by a φ29-encoded, 174-base, RNA and a viral ATPase protein, both of which have fivefold symmetry in three-dimensional cryo-electron microscopy reconstructions. DNA is translocated into the prohead through a 36 Å diameter pore in the center of the connector, where the DNA takes the role of a motor spindle. The helical nature of the DNA allows the rotational action of the connector to be transformed into a linear translation of the DNA. The crystal structure determination of connector crystals in space group C2 was initiated by molecular replacement, using an approximately 20 Å resolution model derived from cryo-electron microscopy. The model phases were extended to 3.5 Å resolution using 12-fold non-crystallographic symmetry averaging and solvent flattening. Although this electron density was not interpretable, the phases were adequate to locate the position of 24 mercury sites of a thimerosal heavy-atom derivative. The resultant 3.2 Å single isomorphous replacement phases were improved using density modification, producing an interpretable electron-density map. The crystallographically refined structure was used as a molecular-replacement model to solve the structures of two other crystal forms of the connector molecule. One of these was in the same space group and almost isomorphous, whereas the other was in space group P21212. The structural differences between the oligomeric connector molecules in the three crystal forms and between different monomers within each crystal show that the structure is relatively flexible, particularly in the protruding domain at the wide end of the connector. This domain probably acts as a bearing, allowing the connector to rotate within the pentagonal portal of the prohead during DNA packaging.

Original languageEnglish (US)
Pages (from-to)1260-1269
Number of pages10
JournalActa Crystallographica Section D: Biological Crystallography
Volume57
Issue number9
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

bacteriophages
Bacteriophages
connectors
Head
Cryoelectron Microscopy
DNA
Crystals
deoxyribonucleic acid
Electron microscopy
Carrier concentration
DNA Packaging
Electrons
Thimerosal
Bearings (structural)
Molecular Models
Viral Proteins
Mercury
Molecules
Adenosine Triphosphatases
crystals

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics
  • Structural Biology

Cite this

Structure determination of the head-tail connector of bacteriophage φ29. / Simpson, A. A.; Leiman, Petr; Tao, Y.; He, Y.; Badasso, M. O.; Jardine, P. J.; Anderson, D. L.; Rossmann, M. G.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 57, No. 9, 2001, p. 1260-1269.

Research output: Contribution to journalArticle

Simpson, AA, Leiman, P, Tao, Y, He, Y, Badasso, MO, Jardine, PJ, Anderson, DL & Rossmann, MG 2001, 'Structure determination of the head-tail connector of bacteriophage φ29', Acta Crystallographica Section D: Biological Crystallography, vol. 57, no. 9, pp. 1260-1269. https://doi.org/10.1107/S0907444901010435
Simpson, A. A. ; Leiman, Petr ; Tao, Y. ; He, Y. ; Badasso, M. O. ; Jardine, P. J. ; Anderson, D. L. ; Rossmann, M. G. / Structure determination of the head-tail connector of bacteriophage φ29. In: Acta Crystallographica Section D: Biological Crystallography. 2001 ; Vol. 57, No. 9. pp. 1260-1269.
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