GYRASE-DEPENDENT INITIATION OF BACTERIOPHAGE-T4 DNA-REPLICATION - INTERACTIONS OF ESCHERICHIA-COLI GYRASE WITH NOVOBIOCIN, COUMERMYCIN AND PHAGE DNA DELAY GENE PRODUCTS

Wild-type bacteriophage T4 and DNA-delay am mutants defective in genes 39, 52, 60 and 58-61 were tested for intracellular sensitivity to the antibiotics coumermycin and novobiocin, drugs which inhibit the DNA gyrase of Escherichia coli. Treatment with these antibiotics drastically reduced the characteristic growth of gene 39, 52 and 60 DNA-delay am mutants in E. coli lacking an amber suppressor (su-). Wild-type phage-infected cells were unaffected by the drugs while the burst size of a gene 58-61 mutant was affected to an intermediate extent. A su-E. coli strain which is resistant to coumermycin due to an altered gyrase permitted growth of the DNA-delay am mutants in the presence of the drug. Thus, the characteristic growth of the DNA-delay am mutants in an su- host apparently depends on the host gyrase. An E. coli himB mutant is defective in the coumermycin-sensitive subunit of gyrase (H. I. Miller, personal communication). Growth of the gene 39, 52 and 60 am mutants was inhibited in the himB mutant while the gene 58-61 mutant and wild-type T4 showed small reductions in burst size in this host. Experiments with nalidixic acid-sensitive and resistant strains of E. coli show that wild-type phage T4 requires a functional nalA protein for growth. Novobiocin and coumermycin inhibit phage DNA synthesis in DNA-delay mutant-infected su-E. coli if added during the early logarithmic phase of phage DNA synthesis. The gene 58-61 mutant showed the smallest inhibition of DNA synthesis in the presence of the drugs. Addition of the drugs during the late linear phase of phage DNA synthesis had no effect on further synthesis in DNA-delay mutant-infected cells. Coumermycin and novobiocin had no effect on DNA synthesis in wild-type-infected cells regardless of the time of addition of the antibiotics. Models are considered in which the DNA-delay gene products either form an autonomous phage gyrase or interact with the host gyrase and adapt it for proper initiation of phage DNA replication. © 1979.