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Moolman, M.C. (author), Tiruvadi Krishnan, S. (author), Kerssemakers, J.W.J. (author), Van den Berg, A. (author), Tulinski, P. (author), Depken, M. (author), Reyes-Lamothe, R. (author), Sherratt, D.J. (author), Dekker, N.H. (author)
The ubiquitous sliding clamp facilitates processivity of the replicative polymerase and acts as a platform to recruit proteins involved in replication, recombination and repair. While the dynamics of the E. coli ?2-sliding clamp have been characterized in vitro, its in vivo stoichiometry and dynamics remain unclear. To probe both ?2-clamp...
journal article 2014
document
Dulin, D. (author), Vilfan, I.D. (author), Berghuis, B.A. (author), Hage, S. (author), Bamford, D.H. (author), Poranen, M.M. (author), Depken, S.M. (author), Dekker, N.H. (author)
RNA viruses have specific mutation rates that balance the conflicting needs of an evolutionary response to host antiviral defenses and avoidance of the error catastrophe. While most mutations are known to originate in replication errors, difficulties of capturing the underlying dynamics have left the mechanochemical basis of viral mutagenesis...
journal article 2015
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Berghuis, B.A. (author), Dulin, D. (author), Xu, Zhi Qiang (author), van Laar, T. (author), Cross, B. (author), Janissen, R. (author), Jergic, Slobodan (author), Dixon, Nicholas E. (author), Depken, S.M. (author), Dekker, N.H. (author)
The bidirectional replication of a circular chromosome by many bacteria necessitates proper termination to avoid the head-on collision of the opposing replisomes. In Escherichia coli, replisome progression beyond the termination site is prevented by Tus proteins bound to asymmetric Ter sites. Structural evidence indicates that strand...
journal article 2015
document
Berghuis, B.A. (author), Raducanu, Vlad-Stefan (author), Elshenawy, Mohamed M. (author), Jergic, Slobodan (author), Depken, S.M. (author), Dixon, NE (author), Hamdan, Samir M. (author), Dekker, N.H. (author)
Synchronizing the convergence of the two-oppositely moving DNA replication machineries at specific termination sites is a tightly coordinated process in bacteria. In Escherichia coli, a “replication fork trap” – found within a chromosomal region where forks are allowed to enter but not leave – is set by the protein–DNA roadblock Tus–Ter. The...
journal article 2017
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Dulin, D. (author), Berghuis, B.A. (author), Depken, S.M. (author), Dekker, N.H. (author)
Single-molecule experiments provide a unique means for real-time observation of the activity of individual biomolecular machines. Through such techniques, insights into the mechanics of for example, polymerases, helicases, and packaging motors have been gleaned. Here we describe the recent advances in single-molecule force spectroscopy...
journal article 2015
document
Janissen, R. (author), Eslami Mossallam, B. (author), Artsimovitch, Irina (author), Depken, S.M. (author), Dekker, N.H. (author)
Pausing by bacterial RNA polymerase (RNAp) is vital in the recruitment of regulatory factors, RNA folding, and coupled translation. While backtracking and intra-structural isomerization have been proposed to trigger pausing, our mechanistic understanding of backtrack-associated pauses and catalytic recovery remains incomplete. Using high...
journal article 2022
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Dulin, D. (author), Arnold, Jamie J. (author), van Laar, T. (author), Oh, Hyung Suk (author), Lee, Cheri (author), Perkins, Angela L. (author), Harki, Daniel A. (author), Depken, S.M. (author), Cameron, Craig E. (author), Dekker, N.H. (author)
RNA viruses pose a threat to public health that is exacerbated by the dearth of antiviral therapeutics. The RNA-dependent RNA polymerase (RdRp) holds promise as a broad-spectrum, therapeutic target because of the conserved nature of the nucleotide-substrate-binding and catalytic sites. Conventional, quantitative, kinetic analysis of antiviral...
journal article 2017
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