Pf1 Filamentous Phage as an Alignment Tool for Generating Local and Global Structural Information in Nucleic Acids

Abstract

Pf1 filamentous phage represent a simple versatile method for generating partially ordered macromolecules in solution. The phage allow tunable degrees of alignment of macromolecules under a wide range of temperature and solvent conditions. The negatively charged phage are ideal for aligning negatively charged nucleic acids and these phage-nucleic acid solutions are stable indefinitely. We have used Pf1 phage to align various DNA and RNA molecules in solution for measurement of dipolar coupling interactions. These dipolar couplings can be used to improve the local structure of nucleic acids. More importantly they also contain information on the global structure, such as DNA bending, which presently cannot be obtained by standard NMR methods. The principles involved in using Pf1 phage to generate solutions of partially order macromolecules will be discussed. The use of ¹H-¹H, ¹H-¹³C and ¹H-¹⁵N dipolar couplings for generating angle constraints for structure refinement of nucleic acids will also be discussed.