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
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Mechanism of substrate translocation by a ring-shaped ATPase motor at millisecond resolution.
Wen Ma,Klaus Schulten +1 more
TL;DR: The present study reveals in new detail the mechanism employed by ring-shaped ATPase motors, for example the use of loosely bound and tightly bound hydrolysis reactant and product states to coordinate motor action, and provides an effective approach to identify allosteric sites of multimeric enzymes in general.
Journal ArticleDOI
Helicase and Polymerase Move Together Close to the Fork Junction and Copy DNA in One-Nucleotide Steps
Manjula Pandey,Smita S. Patel +1 more
TL;DR: This work shows that T7 DNA polymerase and T7 gp4 helicase move in sync during leading-strand synthesis, taking one-nucleotide steps and hydrolyzing one dNTP per base-pair unwound/copied by simultaneously measuring DNA synthesis and d NTP hydrolysis.
Journal ArticleDOI
Single-molecule perspectives on helicase mechanisms and functions.
Bo Sun,Michelle D. Wang +1 more
TL;DR: Single-molecule techniques provide the capability to distinguish helicase heterogeneity and monitor helicase motion at nanometer spatial and millisecond temporal resolutions, ultimately providing new insights into the mechanisms that could not be resolved by ensemble assays.
Journal ArticleDOI
Force-Dependent Stimulation of RNA Unwinding by SARS-CoV-2 NSP13 Helicase
Keith J. Mickolajczyk,Patrick M.M. Shelton,Michael Grasso,Xiaocong Cao,Xiaocong Cao,Sara E. Warrington,Amol Aher,Shixin Liu,Tarun M. Kapoor +8 more
TL;DR: It is found that nsp13 unwinding frequency, processivity, and velocity increase substantially when a destabilizing force is applied to the RNA substrate, and mechanoregulation, which may be provided by a directly bound RNA-dependent RNA polymerase, enables on-demand helicase activity on the relevant polynucleotide substrate during viral replication.
Journal ArticleDOI
Different mechanisms for translocation by monomeric and hexameric helicases
TL;DR: This review surveys structures of monomeric and hexameric helicases and compares different mechanisms for translocation and shows how helicase and polymerase cooperatively catalyze DNA unwinding.
References
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Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche.
Toshiro Sato,Robert G.J. Vries,Hugo J. Snippert,Marc van de Wetering,Nick Barker,Daniel E. Stange,Johan H. van Es,Arie Abo,Pekka Kujala,Peter J. Peters,Hans Clevers +10 more
TL;DR: It is concluded that intestinal cryptvillus units are self-organizing structures, which can be built from a single stem cell in the absence of a non-epithelial cellular niche.
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A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics
TL;DR: Six of the studies are actually in remarkable agreement with one another and explanations are provided in cases where discrepancies remain, and a single set of parameters, derived from 108 oligonucleotide duplexes, adequately describes polymer and oligomer thermodynamics.
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A practical guide to single-molecule FRET
TL;DR: A practical guide to using smFRET, focusing on the study of immobilized molecules that allow measurements of single-molecule reaction trajectories from 1 ms to many minutes, is provided.
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Predicting DNA duplex stability from the base sequence.
TL;DR: This work reports the complete thermodynamic library of all 10 Watson-Crick DNA nearest-neighbor interactions and shows how these thermodynamic data can be used to calculate the stability and predict the temperature-dependent behavior of any DNA duplex structure from knowledge of its base sequence.