Abstract
The presence of RNA-linked nascent DNA pieces in T7 phage-infected Escherichia coli cells has been shown by the selective degradation of the 5'-hydroxyl-terminated nascent DNA, produced by alkali or RNase treatment, with spleen exonuclease. At 43 degrees C, the proportion of RNA-linked DNA pieces in nascent short dna is 50 to 60% in T7 ts136 (ts mutant of gene 6) phage-infected E. coli, whereas that in T7 wild-type phage-infected cells is less than 6%. Joining of the nascent pieces is greatly retarded in T7 ts136-infected E. coli temperature sensitive polA mutants at 43 degrees C. These results suggest that gene 6 exonuclease plays a role in removal of the linked RNA during the discontinuous replication of T7 DNA.
MeSH Terms
Animals
Centrifugation, Density Gradient
Coliphages/metabolism
DNA Replication
DNA, Viral/metabolism
Escherichia coli
Exonucleases/pharmacology
Genes, Viral
Hot Temperature
Mutation
RNA, Viral/metabolism
Swine
Chemicals
DNA, Viral
RNA, Viral
Exonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shinozaki K
Okazaki T
References (20)
20 references, click to expand
-
A mutant of bacteriophage T7 deficient in polynucleotide ligase.
J Biol Chem. 1971 Nov 25;246(22):6874-9
PMID: 5001611
-
The role of bacteriophage T7 exonuclease (gene 6) in genetic recombination and production of concatemers.
J Mol Biol. 1976 Feb 25;101(2):223-34
PMID: 772218
-
Gene 6 exonuclease of bacteriophage T7. II. Mechanism of the reaction.
J Biol Chem. 1972 Jan 10;247(1):311-8
PMID: 4553438
-
Studies on bacteriophage T7 DNA synthesis in vitro. II. Reconstitution of the T7 replication system using purified proteins.
Mol Gen Genet. 1975 Dec 1;141(3):233-49
PMID: 54868
-
Replication of duplex DNA by bacteriophage T7 DNA polymerase and gene 4 protein is accompanied by hydrolysis of nucleoside 5'-triphosphates.
Proc Natl Acad Sci U S A. 1977 Apr;74(4):1525-9
PMID: 323856
-
Mechanism of DNA chain growth XVI. Analyses of RNA-linked DNA pieces in Escherichia coli with polynucleotide kinase.
J Mol Biol. 1977 May 5;112(1):121-40
PMID: 328894
-
Bacteriophage-T7-induced DNA-priming protein. A novel enzyme involved in DNA replication.
Eur J Biochem. 1977 Feb;72(3):543-58
PMID: 320003
-
Degradation of Escherichia coli B deoxyribonucleic acid after infection with deoxyribonucleic acid-defective amber mutants of bacteriophage T7.
J Virol. 1970 Aug;6(2):149-55
PMID: 4925772
-
Deoxyribonucleic acid polymerase of bacteriophage T7.
J Biol Chem. 1971 Nov 25;246(22):6867-73
PMID: 4942327
-
Mechanism of DNA chain growth. XV. RNA-linked nascent DNA pieces in Escherichia coli strains assayed with spleen exonuclease.
J Mol Biol. 1975 Aug 25;96(4):653-64
PMID: 1104864
-
Some functions involved in bacteriophage T7 genetic recombination.
Mol Gen Genet. 1974;134(2):173-80
PMID: 4617162
-
Transient accumulation of Okazaki fragments as a result of uracil incorporation into nascent DNA.
Proc Natl Acad Sci U S A. 1977 Jan;74(1):154-7
PMID: 319455
-
Bacteriophage T7-DNA replication in vitro: an experimental system.
Eur J Biochem. 1973 Sep 21;38(1):160-9
PMID: 4590121
-
Bacteriophage T7.
Science. 1972 Apr 28;176(4033):367-76
PMID: 4554613
-
Evidence that both growing DNA chains at a replication fork are synthesized discontinuously.
Biochemistry. 1976 May 4;15(9):1838-43
PMID: 817735
-
Bacteriophage T7 deoxyribonucleic acid replication in vitro. Requirements for deoxyribonucleic acid synthesis and characterization of the product.
J Biol Chem. 1974 May 10;249(9):2974-80
PMID: 4597463
-
Escherichia coli thioredoxin: a subunit of bacteriophage T7 DNA polymerase.
Proc Natl Acad Sci U S A. 1976 Mar;73(3):780-4
PMID: 768986
-
Formation of concatemeric DNA in bacteriophage T7-infected bacteria.
Virology. 1975 Jun;65(2):455-68
PMID: 1093318
-
Gene 6 exonuclease of bacteriophage T7. I. Purification and properties of the enzyme.
J Biol Chem. 1972 Jan 10;247(1):305-10
PMID: 4336366
-
The genetics and physiology of bacteriophage T7.
Virology. 1969 Nov;39(3):562-74
PMID: 4902069