Crystal structure of an archaeal DNA sliding clamp
clamp molecules from other biological domains shows that the basic mechanism for processive DNA synthesis in DNA replication is conserved in the three domains of life. Load-ing the sliding clamp onto the DNA strand requires the protein factor called the clamp loader. Eukaryotic replica-tion factor C (RFC) (Waga and Stillman 1998), bacterial
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Crystal structures and biochemical characterization of DNA
Crystal structures and biochemical characterization of DNA sliding clamps from three Gram-negative Bacterial sliding clamps bind to DNA and act as protein-protein interaction hubs for several proteins involved Jergic et al., 2013). The clamps are loaded onto dsDNA by a clamp loader complex, comprised of Pol III subunits ( / ) 3
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Clamp and clamp loader structures of the human checkpoint
Background: We have reported that protein imaging by transmission electron microscope observation based on low-angle platinum shadowing can reproduce characteristic ring structures of the replication clamp, proliferating cell nuclear antigen (PCNA), and the clamp loader protein, replication factor C (RFC). The checkpoint protein complexes, Rad9-Hus1-Rad1 (RadLiming) and Rad17-RFCs2-5 (Rad17-RFC
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The Mechanism of ATP-Dependent Primer-Template Recognition
May 15, 2009 · Figures from the paper. Click on the small image to get a bigger one. Figure 1: Structure of the clamp loader:DNA complex (A) Schematic diagram of the clamp loader cycle. (B) Unbiased electron density for the DNA, calculated using a model at a stage prior to the inclusion of DNA and improved using density modification (Terwilliger, 2000).
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(PDF) Biochemical Characterization of DNA Damage
Biochemical Characterization of DNA Damage Checkpoint Complexes: Clamp Loader and Clamp Complexes with Specificity for 5′ Recessed DNA.pdf Available via …
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The β Sliding Clamp Closes around DNA prior to Release by
Escherichia coli γ complex clamp loader functions to load the β sliding clamp onto sites of DNA replication and repair. The clamp loader uses the energy of ATP binding and hydrolysis to drive conformational changes allowing for β binding and opening, DNA binding, and then release of the β·DNA complex. Although much work has been done studying the sliding clamp and clamp loader …
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RCSB PDB - 3U61: Structure of T4 Bacteriophage Clamp
Oct 11, 2011 · The clamp loader traps a spiral conformation of the open clamp so that both the loader and the clamp match the helical symmetry of DNA. One structure reveals that ATP has been hydrolyzed in one subunit and suggests that clamp closure and ejection of the loader involves disruption of the ATP-dependent match in symmetry.
Get a quotePLoS Biochemical Characterization of DNA Damage
Characterization of the biochemical loading reaction executed by the checkpoint clamp loader RSR suggests new insights into the mechanisms underlying recognition of damage-induced DNA structures and signaling to cell cycle controls.
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Minireview DNA polymerase clamp loaders and DNA recognition
Fig. 1. The clamp loading cycle. ATP binding to the clamp loader allows a tight complex to form between clamp loader and sliding clamp. In the ATP-bound state, the clamp loader has a high affinity for primer-template junctions with recessed 30 ends. Binding to these junctions stimulates ATP hydrolysis and release of clamp and DNA,
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Dual Functions, Clamp Opening and Primer-Template
of clamp loader structure and biochemical activity across different species suggests a common overall mechanism of clamp assembly. A key piece of information missing from our knowledge of the clamp loader mechanism is how this multi-protein complex binds DNA and recog-nizes primer-template as the target site for clamp
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Research | Cann Lab at UIUC - University of Illinois at
The M. acetivorans clamp loader is unique and appears to be a "missing link" in the evolution of very complex clamp loaders, as found in humans, from the very simple forms found in P. furiosus. Finally, we have an extensive collaboration with Satish Nair's lab on structural studies of DNA replication proteins and also on enzymes of plant
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Motors and switches: AAA+ machines within the replisome
In particular, studies of the Escherichia coli clamp loader--an AAA+ machine--have provided insights into the architecture of clamp loaders from eukaryotes, bacteriophage T4 and archaea. Other AAA+ proteins are also involved in the initiation of DNA replication, and studies of the E. coli clamp loader indicate mechanisms by which these proteins
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Biochemical characterisation of the clamp/clamp loader
Biochemical characterisation of the clamp/clamp loader proteins from the euryarchaeon Archaeoglobus fulgidus. Nucleic Acids Research, 2002. Sarah Scaife. Martin Singleton. Download PDF. Download Full PDF Package. This paper. A short summary of this paper.
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Replication Clamps and Clamp Loaders
Crystal Structures. Based on early biochemical assays, it had been predicted that sliding clamps encircle DNA in a sequence-independent manner that allows them to slide freely along DNA (Stukenberg et al. 1991).In 1992, this hypothesis was confirmed when the first crystal structure of a sliding clamp, E. coli β clamp, was solved (Fig. 1A, left).
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Clamp loader ATPases and the evolution of DNA replication
Apr 20, 2012 · Clamp loaders are pentameric ATPases of the AAA+ family that operate to ensure processive DNA replication. They do so by loading onto DNA the ring-shaped sliding clamps that tether the polymerase to the DNA. Structural and biochemical analysis of clamp loaders has shown how, despite differences in composition across different branches of life, all clamp loaders undergo the …
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Biochemical characterization of a clamp-loader complex
Aug 04, 2000 · Here we report the isolation and characterization of a clamp-loader complex from the thermoacidophilic archaeon Sulfolobus solfataricus (SsoRFC). SsoRFC is a hetero-pentamer composed of polypeptides of 37 kDa (small subunit) and 46 kDa (large subunit), which possess primary structure similarity with human replication factor C p40 and p140 subunits, respectively.
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Molecular Mechanisms of DNA Polymerase Clamp Loaders
Oct 17, 2011 · Ellison V, Stillman B (2003) Biochemical characterization of DNA damage checkpoint complexes: clamp loader and clamp complexes with specificity for 5′ recessed DNA. PLoS Biol 1:E33 CrossRef Google Scholar
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Replication Clamps and Clamp Loaders
Crystal Structures. Based on early biochemical assays, it had been predicted that sliding clamps encircle DNA in a sequence-independent manner that allows them to slide freely along DNA (Stukenberg et al. 1991).In 1992, this hypothesis was confirmed when the first crystal structure of a sliding clamp, E. coli β clamp, was solved (Fig. 1A, left).
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The clamp-loading complex for processive DNA replication
Jun 20, 2004 · A clamp loader complex loads such clamps onto the DNA strand by opening the clamp ring while at the same time binding and hydrolyzing ATP. All of the sliding clamps, including the Escherichia coli
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Biochemical characterisation of the clamp/clamp loader
Oct 15, 2002 · Abstract. Replicative polymerases of eukaryotes, prokaryotes and archaea obtain processivity using ring‐shaped DNA sliding clamps that are loaded onto DNA by cl
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