In vitro host cell reactivation of alkylated bacteriophage T7 deoxyribonucleic acid by repair-deficient strains of Escherichia coli

Journal Article · Tue Sep 01 04:00:00 EDT 1981 · J. Bacteriol.; (United States)

OSTI ID:6078974

Dodson, L A ^[1]; Masker, W E

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1. Univ. of Tennessee-Oak Ridge Graduate School of Biomedical Sciences, Oak Ridge

An in vitro system capable of packaging bacteriophage T7 deoxyribonucleic acid (DNA) into phage heads to form viable phage particles has been used to monitor the biological consequences of DNA damaged by alkylating agents, and an in vitro DNA replication system has been used to examine the ability of alkylated T7 DNA to serve as template for DNA synthesis. The survival of phage resulting from in vitro packaging of DNA preexposed to various concentrations of methyl methane sulfonate or ethyl methane sulfonate closely paralleled the in vivo situation, in which intact phage were exposed to the alkylating agents. Host factors responsible for survival of alkylated T7 have been examined by using wild-type strains of Escherichia coli and mutants deficient in DNA polymerase I (polA) or 3-methyladeninereverse arrowDNA glycosylase (tag). For both in vivo in vitro situations, a deficiency in 3-methyladenine-DNA glycosylase dramatically reduced phage survival relative to that in the wild type, whereas a deficiency in DNA polymerase I had an intermediate effect. Furthermore, when the tag mutant was used as an indicator strain, phage survival was enhanced when alkylated DNA was packaged with extracts prepared from a wild-type strain in place of the tag mutant or by complementing a tag extract with an uninfected extract, indicating in vitro repair during packaging.

• DOE Contract Number: W-7405-ENG-26
• OSTI ID: 6078974
• Journal Information: J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 147:3; ISSN JOBAA
• Country of Publication: United States
• Language: English

Related Subjects

550700* -- Microbiology
59 BASIC BIOLOGICAL SCIENCES
ALKYLATING AGENTS
BACTERIA
BACTERIOPHAGES
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CELL CULTURES
DNA
DNA-ASE
ENZYMES
ESCHERICHIA COLI
ESTERASES
GLYCOSYL HYDROLASES
HOST-CELL REACTIVATION
HYDROLASES
IN VITRO
MICROORGANISMS
MONITORING
MUTANTS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PARASITES
PHOSPHODIESTERASES
RECOVERY
REPAIR
VIRUSES