Relationship of Bacillus subtilis DNA polymerase III to bacteriophage PBS2-induced DNA polymerase and to the replication of uracil-containing DNA
In vivo studies of PBS2 phage replication in a temperature-sensitive Bacillus subtilis DNA polymerase III (Pol III) mutant and a temperature-resistant revertant of this mutant have suggested the possible involvement of Pol III in PBS2 DNA synthesis. Previous results with 6-(p-hydroxyphenylazo)-uracil (HPUra), a specific inhibitor of Pol III and DNA replication in uninfected cells, suggest that Pol III is not involved in phage DNA replication, due to its resistance to this drug. Experiments were designed to examine possible explanations for this apparent contradiction. First, assays of the host Pol III and the phage-induced DNA polymerase activities in extracts indicated that a labile Pol III did not result in a labile phage-induced enzyme, suggesting that this new polymerase is not a modified HPUra-reisistant form of Pol III. Indeed the purified phage-induced enzyme was resistant to the active, reduced form of HPUra under all assay conditions tested. Since in vitro Pol III was capable of replicating the uracil-containing DNA found in this phage, the sensitivity of the purified enzyme to reduced HPUra was examined using phage DNA as template-primer and dUTP as substrate; these new substrates did not affect the sensitivity of the host enzyme to the drug.
- Research Organization:
- Univ. of Michigan, Ann Arbor
- OSTI ID:
- 6531927
- Journal Information:
- J. Virol.; (United States), Vol. 28:3
- Country of Publication:
- United States
- Language:
- English
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Synthesis of DNA containing uracil during bacteriophage infection of Bacillus subtilis. Technical progress report, August 1, 1975--July 31, 1976
Related Subjects
BACILLUS SUBTILIS
BIOCHEMICAL REACTION KINETICS
BACTERIOPHAGES
DNA
DNA REPLICATION
POLYMERASES
METABOLISM
MUTANTS
URACILS
AZINES
BACILLUS
BACTERIA
ENZYMES
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
KINETICS
MICROORGANISMS
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PYRIMIDINES
REACTION KINETICS
VIRUSES
550200* - Biochemistry
550700 - Microbiology