Abstract
Experiments with a mutant of T4, tsL97, temperature sensitive for gene 43, showed that T4 DNA polymerase was necessary in vivo to repair gaps in recombinant molecules. CsCl density gradient experiments showed that molecular recombinants were not repaired when the T4tsL97-infected cells were shifted to 42 C after replication and recombination had taken place. Repair was almost complete when the same procedure was followed with the wild-type T4, or when the T4tsL97-infected cells were incubated at the permissive temperature, 36 C. Long-single-strand production was also affected similarly by the T4tsL97 mutation. All the results were consistent with the theory that gaps exist in many recombinant molecules at the recombinant joint, that T4 DNA polymerase is the enzyme that repairs these gaps in vivo, and that covalent repair of the recombinants leads to extensive long-single-strand production.
MeSH Terms
Centrifugation, Density Gradient
Coliphages/enzymology,metabolism
DNA Nucleotidyltransferases/biosynthesis,metabolism
DNA Repair
DNA Viruses
DNA, Single-Stranded/biosynthesis
DNA, Viral/biosynthesis
Genes
Mutation
Phosphorus Radioisotopes
Recombination, Genetic
Temperature
Chemicals
DNA, Single-Stranded
DNA, Viral
Phosphorus Radioisotopes
DNA Nucleotidyltransferases
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Miller R C
References (22)
22 references, click to expand
-
Early intracellular events in the replication of T4 phage DNA. I. Complex formation of replicative DNA.
Proc Natl Acad Sci U S A. 1965 Jul;54(1):273-8
PMID: 5323019
-
Evidence for long DNA strands in the replicating pool after T4 infection.
Proc Natl Acad Sci U S A. 1968 Jan;59(1):131-8
PMID: 4873341
-
Early intracellular events in the replication T4 phage DNA. II. Partially replicated DNA.
Proc Natl Acad Sci U S A. 1965 Aug;54(2):634-40
PMID: 5218656
-
Molecular recombination in T4 bacteriophage deoxyribonucleic acid. I. Tertiary structure of early replicative and recombining deoxyribonucleic acid.
J Virol. 1967 Aug;1(4):758-70
PMID: 5621474
-
Molecular Recombination in T4 Bacteriophage Deoxyribonucleic Acid: III. Formation of Long Single Strands During Recombination.
J Virol. 1970 Mar;5(3):368-80
PMID: 16789127
-
Dispersive transfer of the parental DNA molecule to the progeny of phage phiX-174.
Virology. 1959 Oct;9:260-74
PMID: 14411537
-
REPLICATIVE FRAGMENTATION IN T4 BACTERIOPHAGE DNA. II. BIPARENTAL MOLECULAR RECOMBINATION.
Proc Natl Acad Sci U S A. 1964 Aug;52:211-8
PMID: 14206583
-
THE SYNTHESIS OF MESSENGER RNA WITHOUT PROTEIN SYNTHESIS. II. SYNTHESIS OF PHAGE-INDUCED RNA AND SEQUENTIAL ENZYME PRODUCTION.
J Mol Biol. 1964 May;8:638-59
PMID: 14187391
-
MOLECULAR MECHANISMS OF GENETIC RECOMBINATION IN BACTERIOPHAGE. II. JOINING OF PARENTAL DNA MOLECULES OF PHAGE T4.
J Mol Biol. 1964 Apr;8:516-40
PMID: 14153524
-
Fragmentary transfer of P32-labeled parental DNA to progeny phage.
Virology. 1961 Jan;13:124-34
PMID: 13753771
-
Suppression of amber mutations of bacteriophage T4 gene 43 (DNA polymerase) by translational ambiguity.
J Virol. 1973 Jun;11(6):933-45
PMID: 4351461
-
Molecular mechanisms of genetic recombination of bacteriophage. V. Two kinds of joining of parental DNA molecules.
J Mol Biol. 1965 Jul;12(3):805-15
PMID: 4221791
-
T7 exonuclease (gene 6) is necessary for molecular recombination of bacteriophage T7.
J Virol. 1974 Nov;14(5):1040-8
PMID: 4610173
-
Replication of T4 DNA in Escherichia coli treated with toluene.
J Virol. 1973 Dec;12(6):1195-203
PMID: 4586770
-
An electron microscopic analysis of pathways for bacteriophage T4 DNA recombination.
J Mol Biol. 1973 Nov 25;81(1):1-16
PMID: 4588574
-
Role of genes 46 and 47 in bacteriophage T4 reproduction. II. Formation of gaps on parental DNA of polynucleotide ligase defective mutants.
J Mol Biol. 1972 Oct 14;70(3):617-35
PMID: 4563264
-
Branched DNA molecules: intermediates in T4 recombination.
J Mol Biol. 1971 Aug 28;60(1):131-49
PMID: 4937189
-
Molecular recombination in T4 bacteriophage deoxyribonucleic acid. II. Single-strand breaks and exposure of uncomplemented areas as a prerequisite for recombination.
J Virol. 1967 Dec;1(6):1193-202
PMID: 4912244
-
Enzymic joining of polynucleotides. VII. Role of the T4-induced ligase in the formation of recombinant molecules.
J Mol Biol. 1969 Dec 28;46(3):467-79
PMID: 4904103
-
Enzymatic synthesis of deoxyribonucleic acid. XXXII. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break.
J Biol Chem. 1970 Jan 10;245(1):39-45
PMID: 4904090
-
Enzymic synthesis of DNA. 33. Hydrolysis of a 5'-triphosphate-terminated polynucleotide in the active center of DNA polymerase.
J Mol Biol. 1969 Nov 14;45(3):513-31
PMID: 4917367
-
The structural gene for deoxyribonucleic acid polymerase in bacteriophages T4 and T5.
Proc Natl Acad Sci U S A. 1965 Oct;54(4):1241-8
PMID: 5219829