TY - JOUR
T1 - Coordinated leading and lagging strand DNA synthesis on a minicircular template
AU - Lee, Joonsoo
AU - Chastain, Paul D.
AU - Kusakabe, Takahiro
AU - Griffith, Jack D.
AU - Richardson, Charles C.
N1 - Funding Information:
We are grateful to Stan Tabor for suggesting the use of a minicircle for these studies. We thank Tom Ellenberger and Ingrid Richardson for critical comments. This work was supported by grants to C. C. R. from the National Institutes of Health (AI-06045) and the American Cancer Society (NP-I-28), and to J. D. G. from the National Institutes of Health (GM31819). C. C. R. is a consultant to Amersham Life Science Inc., which has licenses from Harvard University to commercialize DNA replication proteins.
PY - 1998/6
Y1 - 1998/6
N2 - The coordinated synthesis of both leading and lagging DNA strands is thought to involve a dimeric DNA polymerase and a looping of the lagging strand so that both strands can be synthesized in the same direction. We have constructed a minicircle with a replication fork that permits an assessment of the stoichiometry of the proteins and a measurement of the synthesis of each strand. The replisome consisting of bacteriophage T7 DNA polymerase, helicase, primase, and single-stranded DNA-binding protein mediates coordinated replication. The criteria for coordination are fulfilled : (1) a replication loop is formed, (2) leading and lagging strand synthesis are coupled, (3) the lagging strand polymerase recycles from one Okazaki fragment to another, and (4) the length of Okazaki fragments is regulated. T7 single-stranded DNA-binding protein is essential for coordination.
AB - The coordinated synthesis of both leading and lagging DNA strands is thought to involve a dimeric DNA polymerase and a looping of the lagging strand so that both strands can be synthesized in the same direction. We have constructed a minicircle with a replication fork that permits an assessment of the stoichiometry of the proteins and a measurement of the synthesis of each strand. The replisome consisting of bacteriophage T7 DNA polymerase, helicase, primase, and single-stranded DNA-binding protein mediates coordinated replication. The criteria for coordination are fulfilled : (1) a replication loop is formed, (2) leading and lagging strand synthesis are coupled, (3) the lagging strand polymerase recycles from one Okazaki fragment to another, and (4) the length of Okazaki fragments is regulated. T7 single-stranded DNA-binding protein is essential for coordination.
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U2 - 10.1016/S1097-2765(00)80100-8
DO - 10.1016/S1097-2765(00)80100-8
M3 - Article
C2 - 9651583
AN - SCOPUS:0032086491
SN - 1097-2765
VL - 1
SP - 1001
EP - 1010
JO - Molecular Cell
JF - Molecular Cell
IS - 7
ER -