A DNA Gyrase-Binding Site at the Center of the Bacteriophage Mu Genome is Required for Efficient Replicative Transposition

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 87; no. 22; pp. 8716 - 8720
• Main Authors: Pato, Martin L., Howe, Martha M., Higgins, N. Patrick
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01-11-1990; National Acad Sciences
• Subjects: Bacteriophage mu - genetics; Base Sequence; Binding Sites; Biological and medical sciences; DNA; DNA Replication; DNA Topoisomerases, Type II - metabolism; DNA Transposable Elements; DNA, Superhelical - physiology; DNA, Viral - metabolism; Enzymes; Escherichia coli; Fundamental and applied biological sciences. Psychology; Gels; Genetic mutation; Genetic transposition; Genic rearrangement. Recombination. Transposable element; Genomes; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Molecules; Mutation; Plasmids; Prophages; Restriction Mapping; Sequencing; Virus Replication; DNA topoisomerase (ATP-hydrolysing); Transposition; Molecular interaction; Enzyme
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.87.22.8716

We have discovered a centrally located site that is required for efficient replication of bacteriophage Mu DNA and identified it as a strong DNA gyrase-binding site. Incubation of Mu DNA with gyrase and enoxacin revealed a cleavage site 18.1 kilobases from the left end of the 37.2-kilobase genome. Two observations indicate a role for the site in Mu replication: mutants of Mu, able to grow on an Escherichia coli gyrB host that does not allow growth of wild-type Mu, were found to possess single-base changes resulting in more efficient gyrase binding and cleavage at the site. Introduction of a 147-base-pair deletion that eliminated the site from a prophage inhibited the onset of Mu replication for >1 hr after induction.