TY - JOUR
T1 - Enzymic synthesis of deoxyribonucleic acid. XXII. Replication of a circular single-stranded DNA template by DNA polymerase of Escherichia coli
AU - Mitra, Sankar
AU - Reichard, Peter
AU - Inman, R. B.
AU - Bertsch, Le Roy L.
AU - Kornberg, Arthur
N1 - Funding Information:
This investigation was supported in part by grants from the U.S. National Institutes of Health, the National Science Foundation and the National Aeronautics and Space Administration. We acknowledge the assistance of Miss Josephine Van Pinxteren in the electron microscopic studies reported.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1967/3/28
Y1 - 1967/3/28
N2 - DNA from the bacteriophage M13, specific for male strains of Escherichia coli, was found to have a ring structure similar to that of φX174 DNA. E. coli DNA polymerase, free of endonuclease, uses this DNA as a template for a complete or even more extensive replication. The template-product complex is separable in alkali, yielding the original circular template and an open-chain product strand. The molecular weights of the template, the full replication product and the 1:1 hybrid of template and product were calculated from sedimentation coefficients to be 2 × 106, 2 × 106 and 4 × 106 daltons, respectively. Electron microscopic studies of the hybrid reveal circular, helical molecules about 2 μ in contour length; with only partial replication, circular molecules are seen which appear to have one single-stranded region linking the ends of a helical segment; with extensive replication, helical branches are seen attached to 2-μ long helical circles. Analysis of the product reveals the initiating termini of the strands as the 5′-(mono)-phosphates of each of the four deoxynucleotides. Replication of circular M13 DNA thus appears to be initiated without covalent linkage to the template, without a specific nucleotide locus on the template and with the elimination of the terminal pyrophosphoryl group of the initial deoxynucleoside triphosphate.
AB - DNA from the bacteriophage M13, specific for male strains of Escherichia coli, was found to have a ring structure similar to that of φX174 DNA. E. coli DNA polymerase, free of endonuclease, uses this DNA as a template for a complete or even more extensive replication. The template-product complex is separable in alkali, yielding the original circular template and an open-chain product strand. The molecular weights of the template, the full replication product and the 1:1 hybrid of template and product were calculated from sedimentation coefficients to be 2 × 106, 2 × 106 and 4 × 106 daltons, respectively. Electron microscopic studies of the hybrid reveal circular, helical molecules about 2 μ in contour length; with only partial replication, circular molecules are seen which appear to have one single-stranded region linking the ends of a helical segment; with extensive replication, helical branches are seen attached to 2-μ long helical circles. Analysis of the product reveals the initiating termini of the strands as the 5′-(mono)-phosphates of each of the four deoxynucleotides. Replication of circular M13 DNA thus appears to be initiated without covalent linkage to the template, without a specific nucleotide locus on the template and with the elimination of the terminal pyrophosphoryl group of the initial deoxynucleoside triphosphate.
UR - http://www.scopus.com/inward/record.url?scp=49949148266&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=49949148266&partnerID=8YFLogxK
U2 - 10.1016/0022-2836(67)90229-x
DO - 10.1016/0022-2836(67)90229-x
M3 - Article
AN - SCOPUS:49949148266
SN - 0022-2836
VL - 24
SP - 429-442,IN13-IN16,443-447
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 3
ER -