Single-Molecule Fluorescence Reveals the Unwinding Stepping Mechanism of Replicative Helicase
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TLDR
A coupling ratio of 1:1 between base pairs unwound and dTTP hydrolysis is suggested, which further support the concept that nucleic acid motors can have a hierarchy of different-sized steps or can accumulate elastic energy before transitioning to a subsequent phase.About:
This article is published in Cell Reports.The article was published on 2014-03-27 and is currently open access. It has received 54 citations till now. The article focuses on the topics: Helicase.read more
Citations
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Journal ArticleDOI
Open-ringed structure of the Cdt1-Mcm2-7 complex as a precursor of the MCM double hexamer
Yuanliang Zhai,Erchao Cheng,Hao Wu,Ningning Li,Philip Y. K. Yung,Ning Gao,Bik Kwoon Tye,Bik Kwoon Tye +7 more
TL;DR: The intrinsic coiled structures of the precursors provide insights into the DH formation, and suggest a spring-action model for the MCM during the initial origin melting and the subsequent DNA unwinding.
Journal ArticleDOI
Simultaneous Real-Time Imaging of Leading and Lagging Strand Synthesis Reveals the Coordination Dynamics of Single Replisomes
Karl E. Duderstadt,Karl E. Duderstadt,Karl E. Duderstadt,Hylkje Geertsema,Sarah A. Stratmann,Christiaan M. Punter,Arkadiusz W. Kulczyk,Charles C. Richardson,Antoine M. van Oijen,Antoine M. van Oijen +9 more
TL;DR: It is shown that loops in the lagging strand predominantly occur during priming and only infrequently support subsequent Okazaki-fragment synthesis, reconciling divergent models for the regulation of primer synthesis and revealing an underlying plasticity in replisome operation.
Journal ArticleDOI
Single-Molecule Analysis and Engineering of DNA Motors
TL;DR: Techniques such as mutagenesis, chemical modifications, and optogenetics that have been used to re-engineer existing molecular motors to have, for instance, altered speed, processivity, or functionality are reviewed.
Journal ArticleDOI
Mechanical Operation and Intersubunit Coordination of Ring-Shaped Molecular Motors: Insights from Single-Molecule Studies
TL;DR: The various mechanisms by which ring motors convert chemical energy to mechanical force or torque and coordinate the activities of individual subunits that constitute the ring are described.
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The mechanism of DNA unwinding by the eukaryotic replicative helicase.
TL;DR: A mechanism in which purified recombinant Drosophila melanogaster CMG couples ATP hydrolysis to unwinding by acting as a lazy Brownian ratchet is proposed, providing quantitative understanding of the central process in eukaryotic DNA replication.
References
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Oligomeric structure of bacteriophage T7 DNA primase/helicase proteins.
TL;DR: The oligomeric structure of bacteriophage T7 gene 4 helicase/primase proteins was investigated using protein cross-linking and high pressure gel-filtration chromatography and indicated that hexamers were the predominant species to accumulate in both 4A' and 4B proteins.
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A Hexameric Helicase Encircles One DNA Strand and Excludes the Other during DNA Unwinding
TL;DR: It is unambiguously demonstrated that the T7 gene 4 protein is a 5'-to-3' helicase and imply that during DNA unwinding the 5'- to- 3' strand transverses the center of the ring while the 3'-to the 5-to-5' strand is excluded from the centerof the ring.
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Does single-stranded DNA pass through the inner channel of the protein hexamer in the complex with the Escherichia coli DnaB Helicase? Fluorescence energy transfer studies.
TL;DR: Evidence that, in the complex with the enzyme, ssDNA passes through the inner channel of the DnaB hexamer is provided, the first evidence of the existence of such a structure of a hexameric helicase-ssDNA complex in solution.
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Asymmetric interactions of hexameric bacteriophage T7 DNA helicase with the 5'- and 3'-tails of the forked DNA substrate.
Peter Ahnert,Smita S. Patel +1 more
TL;DR: An exclusion model of DNA unwinding is proposed in which T7 helicase hexamer interacts mainly with the ssDNA strands duringDNA unwinding, encircling the 5′-strand and excluding the 3′-Strand from the hole.
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Translocation step size and mechanism of the RecBC DNA helicase.
TL;DR: It is proposed that the RecBC enzyme both translocates and unwinds by a quantized, two-step, inchworm-like mechanism that may have parallels for translocation by other linear motor proteins.