Structure of the head-tail connector of bacteriophage φ29

Carrascosa, J.L.; Viñuela, Eladio; García, Narciso; Santisteban, Antonio

doi:10.1016/0022-2836(82)90066-3

Cited by 64 publications

(41 citation statements)

References 30 publications

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“…Early results demonstrated that 12 appendages are attached to the lower collar and function as adsorption organelles in the mature phage (Anderson et al, 1966;Tosi and Anderson, 1973;Carrascosa et al, 1982). However, the 5-fold averaging inherent in the reconstruction process (Experimental Procedures) produces 10, rather than 12, large density features in a "miniskirt" attached to the bulge of the lower collar ( Figures 2I, 2J, 2N, and 2O).…”

Section: The Neck/tail Assemblymentioning

confidence: 94%

Assembly of a Tailed Bacterial Virus and Its Genome Release Studied in Three Dimensions

Tao¹,

Olson²,

Wang³

et al. 2014

Selected Papers of Michael G Rossmann With Commentaries

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SummaryWe present the first three-dimensional reconstruction of a prolate, tailed phage, and its empty prohead precursor by cryo-electron microscopy. The head-tail connector, the central component of the DNA packaging machine, is visualized for the first time in situ within the Bacillus subtilis dsDNA phage ϕ29. The connector, with 12-or 13-fold symmetry, appears to fit loosely into a pentameric vertex of the head, a symmetry mismatch that may be required to rotate the connector to package DNA. The prolate head of ϕ29 has 10 hexameric units in its cylindrical equatorial region, and 11 pentameric and 20 hexameric units comprise icosahedral end-caps with T=3 quasisymmetry. Reconstruction of an emptied phage particle shows that the connector and neck/tail assembly undergo significant conformational changes upon ejection of DNA.

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Section: The Neck/tail Assemblymentioning

confidence: 94%

Assembly of a Tailed Bacterial Virus and Its Genome Release Studied in Three Dimensions

Tao¹,

Olson²,

Wang³

et al. 2014

Selected Papers of Michael G Rossmann With Commentaries

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show abstract

“…The 12 appendages had been shown to attach to the lower collar and to function as adsorption organelles in the mature phage (Anderson et al, 1966;Tosi and Anderson, 1973;Carrascosa et al, 1982). In the micrographs of the mature phage, the appendages ap- pear as long, spindly objects emanating from the head-tail junction.…”

Section: Asymmetric Reconstruction Of Mature Phage Particlesmentioning

confidence: 95%

Cryoelectron-Microscopy Image Reconstruction of Symmetry Mismatches in Bacteriophage φ29

Morais

Tao

Olson

et al. 2001

Journal of Structural Biology

103

110

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A method has been developed for three-dimensional image reconstruction of symmetry-mismatched components in tailed phages. Although the method described here addresses the specific case where differing symmetry axes are coincident, the method is more generally applicable, for instance, to the reconstruction of images of viral particles that deviate from icosahedral symmetry. Particles are initially oriented according to their dominant symmetry, thus reducing the search space for determining the orientation of the less dominant, symmetry-mismatched component. This procedure produced an improved reconstruction of the sixfold-symmetric tail assembly that is attached to the fivefold-symmetric prolate head of phi29, demonstrating that this method is capable of detecting and reconstructing an object that included a symmetry mismatch. A reconstruction of phi29 prohead particles using the methods described here establishes that the pRNA molecule has fivefold symmetry when attached to the prohead, consistent with its proposed role as a component of the stator in the phi29 DNA packaging motor.

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“…By 3D reconstruction a central channel has been observed (Carazo et al, 1985) that is probably required for translocating the DNA (Carrascosa et al, 1990). Connectors exhibit 12-fold (Driedonks et al, 1981;Carrascosa et al, 1982;Kochan et al, 1984;Carazo et al, 1986;Kocsis et al, 1995;Cerritelli and Studier, 1996) as well as 13-fold (Dube et al, 1993;Tsuprun et al, 1994) rotational symmetry. This interesting polymorphism fostered different models for the DNA The adsorption buffer may vary from 4 to 10 at KCl or NaCl concentrations between 50 and 500 mM; the ideal imaging buffer is pH 8.2-9.2, 10 mM Tris-HCl, 150 mM KCl.…”

Section: Fig 10mentioning

confidence: 99%

Adsorption of Biological Molecules to a Solid Support for Scanning Probe Microscopy

Müller

Amrein

Engel

1997

Journal of Structural Biology

286

204

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Structure of the head-tail connector of bacteriophage φ29

Cited by 64 publications

References 30 publications

Assembly of a Tailed Bacterial Virus and Its Genome Release Studied in Three Dimensions

Assembly of a Tailed Bacterial Virus and Its Genome Release Studied in Three Dimensions

Cryoelectron-Microscopy Image Reconstruction of Symmetry Mismatches in Bacteriophage φ29

Adsorption of Biological Molecules to a Solid Support for Scanning Probe Microscopy

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