Matching Electrostatic Charge between DNA and Coat Protein in Filamentous Bacteriophage. Fibre Diffraction of Charge-deletion Mutants

• Published in: Journal of molecular biology Vol. 245; no. 2; pp. 86 - 91
• Main Authors: Symmons, Martyn F., Welsh, Liam C., Nave, Colin, Marvin, Don A., Perham, Richard N.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 13.01.1995
• Subjects: Capsid - chemistry; DNA, Viral - chemistry; DNA/protein interactios; Electrochemistry; fibre diffraction; Inovirus; Inovirus - chemistry; Inovirus - genetics; macromolcular assemblies; molecular engineering; Mutation; Nucleic Acid Conformation; phage f1; phage fd; phage M13; Protein Conformation; X-Ray Diffraction; molecular engineering; DNA/protein interactios; macromolcular assemblies; Inovirus; fibre diffraction
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1006/jmbi.1994.0009

The virion of Ff (fd, f1, M13) filamentous bacteriophage consists of a long tube of coat protein subunits in a shingled, helical array, surrounding a genome of circular single-stranded DNA. Modified fd virions have been generated by a mutation (K48a) that removes one positive charge from each coat protein subunit in the C-terminal region of the polypeptide chain facing the DNA. The number of nucleotides in the mutant DNA is unchanged, but the K48A virons are 35% longer than wild-type. We have measured the X-ray diffraction attributable to single virions in hydrated gels of wild-type. We have measured the X-ray diffraction attributable to single virons in hydrated gels of wild-type and K48A bacteriophages. Most of the diffraction pattern shows no significant difference between wild-type and K48A. Since the DNA is only about 12% by weight of the wild-type virion, the diffraction pattern is dominated by the protein contribution, and the absence of significant differences indicates that there are no significant changes in the symmetry or structure of the protein coat. But there is a change in the diffraction pattern in a region where the DNA and protein contributions are comparable. The diffraction pattern is a region where the DNA and protein contributions are comparable. The diffraction pattern of the K48A mutant shows an increase in intensity of one of the weaker equatorial peaks, relative to wild-type, in a region where the protein contribution has negative sign but the DNA contribution has positive sign. This is consistent with a decrease in the ratio of DNA:protein per unit length of the K48A mutant. The results support the view that the protein forms a sheath lined with positive charges interacting electrostatically and non-specifically with a negatively charged DNA core of matching charge density. The lower positive charge density lining the capsid in the K48A mutant means that correspondingly fewer nucleotides can be packaged per coat protein subunit, which in turn requires an elongation of the DNA inside the virion. A longer virion is thus required to package the same amount of DNA. Within the error of masurement, the number of positive charges on the protein interacting with the DNA is the same in K48A as in the wild-type, despite the fact that the mutant is 35% longer than the wild-type.