DNA synthesis provides the driving force to accelerate DNA unwinding by a helicase
- 19 May 2005
- journal article
- research article
- Published by Springer Nature in Nature
- Vol. 435 (7040) , 370-373
- https://doi.org/10.1038/nature03615
Abstract
Helicases are molecular motors that use the energy of nucleoside 5′-triphosphate (NTP) hydrolysis to translocate along a nucleic acid strand and catalyse reactions such as DNA unwinding. The ring-shaped helicase1 of bacteriophage T7 translocates along single-stranded (ss)DNA at a speed of 130 bases per second2; however, T7 helicase slows down nearly tenfold when unwinding the strands of duplex DNA3. Here, we report that T7 DNA polymerase, which is unable to catalyse strand displacement DNA synthesis by itself, can increase the unwinding rate to 114 base pairs per second, bringing the helicase up to similar speeds compared to its translocation along ssDNA. The helicase rate of stimulation depends upon the DNA synthesis rate and does not rely on specific interactions between T7 DNA polymerase and the carboxy-terminal residues of T7 helicase. Efficient duplex DNA synthesis is achieved only by the combined action of the helicase and polymerase. The strand displacement DNA synthesis by the DNA polymerase depends on the unwinding activity of the helicase, which provides ssDNA template. The rapid trapping of the ssDNA bases by the DNA synthesis activity of the polymerase in turn drives the helicase to move forward through duplex DNA at speeds similar to those observed along ssDNA.Keywords
This publication has 26 references indexed in Scilit:
- General Methods for Analysis of Sequential “n-step” Kinetic Mechanisms: Application to Single Turnover Kinetics of Helicase-Catalyzed DNA UnwindingBiophysical Journal, 2003
- ATP Binding Modulates the Nucleic Acid Affinity of Hepatitis C Virus HelicasePublished by Elsevier ,2003
- A General Model for Nucleic Acid Helicases and Their “Coupling” within Macromolecular MachinesCell, 2001
- Structure and Function of Hexameric HelicasesAnnual Review of Biochemistry, 2000
- Kinetic Measurement of the Step Size of DNA Unwinding by Escherichia coli UvrD HelicaseScience, 1997
- Processivity of the Gene 41 DNA Helicase at the Bacteriophage T4 DNA Replication ForkJournal of Biological Chemistry, 1996
- ESCHERICHIA COLI SINGLE-STRANDED DNA-BINDING PROTEIN: Multiple DNA-Binding Modes and CooperativitiesAnnual Review of Biochemistry, 1994
- Construction and characterization of a bacteriophage T4 DNA polymerase deficient in 3'-->5' exonuclease activity.Proceedings of the National Academy of Sciences, 1993
- Pre-steady-state kinetic analysis of processive DNA replication including complete characterization of an exonuclease-deficient mutantBiochemistry, 1991
- Bacteriophage T7: Minimal requirements for the replication of a duplex DNA moleculeCell, 1983