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Chapter 59 : Morphogenesis of Bacteriophage ϕ29

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Morphogenesis of Bacteriophage ϕ29, Page 1 of 2

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Abstract:

Bacteriophages are very useful models for studying the protein-derived principles of form determination in virus assembly. Bacteriophage models cannot be ignored because of the assembly of the virus tail at the prohead portal vertex or connector, the entry point of double-stranded DNA. Bacteriophage ø29 of , illustrated in the family portrait with bacteriophage T2 of , features simplicity, efficient in vitro assembly, and advanced genetic and biochemical characterizations. This chapter emphasizes ø29 structure and the mechanisms of prohead assembly and DNA packaging, the major thrusts of current ø29 research in Minneapolis. The basic problem of ø29 morphogenesis is the mechanism by which a prolate shell of particular dimensions is assembled from subunits that are also capable of being assembled into incorrect structures. Mutants of asporogenous that cannot assemble ø29 (vam) were selected by the use of antibodies that reacted more strongly with the free connectors than with the portal vertex of proheads or phage. Bacteriophage DNA-packaging machines seem to have in common two packaging proteins and a connector with sixfold symmetry. Restriction enzyme digestion of DNA molecules extracted from DNase I-treated proheads shows that DNA-gp3 packaging is oriented with respect to the physical map. A gyrase action would be needed to generate supercoiled DNA for packaging.

Citation: Anderson D, Reilly B. 1993. Morphogenesis of Bacteriophage ϕ29, p 859-867. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch59

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Transmission Electron Microscopy
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Scanning Electron Microscopy
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Agarose Gel Electrophoresis
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DNA Restriction Enzymes
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Figures

Image of Figure 1
Figure 1

Family portrait of phages 29 and T2. Magnification, x 428,000.

Citation: Anderson D, Reilly B. 1993. Morphogenesis of Bacteriophage ϕ29, p 859-867. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch59
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Image of Figure 2
Figure 2

Pathway of 29 morphogenesis. Proteins composing a structure are listed below it. Prohead II and the 1¯, 12¯, and 13¯ particles are true intermediates in the pathway. The 10¯, 7¯, and 9¯ structures shown below the main pathway are abortive structures. Particle a has been observed only in restrictive 12(716) infection together with the 12¯ particle; its composition is inferred from morphology.

Citation: Anderson D, Reilly B. 1993. Morphogenesis of Bacteriophage ϕ29, p 859-867. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch59
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Image of Figure 3
Figure 3

Three views of the modified ? = 3 model of the 29 prohead according to S. Casjens and R. Hendrix. The model contains 30 hexamers (light) and 11 pentamers (dark) of the major capsid protein. The panel on the right shows a view of the base, where the connector (not shown) is attached. The computer-generated schematics are by E. Egelman.

Citation: Anderson D, Reilly B. 1993. Morphogenesis of Bacteriophage ϕ29, p 859-867. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch59
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Image of Figure 4
Figure 4

Secondary structure of 29 pRNA derived by phylogenetic analysis ( ). A through F, helical stems. Residues conserved in the 29-???, M2-Nf, SF5, and GA1 pRNA sequences are designated in boldface letters.

Citation: Anderson D, Reilly B. 1993. Morphogenesis of Bacteriophage ϕ29, p 859-867. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch59
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