The decatenation checkpoint

The decatenation checkpoint delays entry into mitosis until the chromosomes have been disentangled. Deficiency in or bypass of the decatenation checkpoint can cause chromosome breakage and nondisjunction during mitosis, which results in aneuploidy and chromosome rearrangements in the daughter cells. A deficiency in the decatenation checkpoint has been reported in lung and bladder cancer cell lines and may contribute to the accumulation of chromosome aberrations that commonly occur during tumour progression. A checkpoint deficiency has also been documented in cultured stem and progenitor cells, and cancer stem cells are likely to be derived from stem and progenitor cells that lack an effective decatenation checkpoint. An inefficient decatenation checkpoint is likely to be a source of the chromosome aberrations that are common features of most tumours, but an inefficient decatenation checkpoint in cancer stem cells could also provide a potential target for chemotherapy

Integrating Ion Mobility Mass Spectrometry with Molecular Modelling to Determine the Architecture of Multiprotein Complexes

Current challenges in the field of structural genomics point to the need for new tools and technologies for obtaining structures of macromolecular protein complexes. Here, we present an integrative computational method that uses molecular modelling, ion mobility-mass spectrometry (IM-MS) and incomplete atomic structures, usually from X-ray crystallography, to generate models of the subunit architecture of protein complexes. We begin by analyzing protein complexes using IM-MS, and by taking measurements of both intact complexes and sub-complexes that are generated in solution. We then examine available high resolution structural data and use a suite of computational methods to account for missing residues at the subunit and/or domain level. High-order complexes and sub-complexes are then constructed that conform to distance and connectivity constraints imposed by IM-MS data. We illustrate our method by applying it to multimeric protein complexes within the Escherichia coli replisome: the sliding clamp, (β2), the γ complex (γ3δδ′), the DnaB helicase (DnaB6) and the Single-Stranded Binding Protein (SSB4)

Phase Transformations from Quartz to Cristobalite

Two quartz types used in the silicon and ferrosilicon industry were heated to temperatures of 1600 and 1700 °C. The parameters varied were the temperature and the holding time at maximum temperature. The amount of quartz, cristobalite and intermediate amorphous phase were measured using XRD and the internal standard method. Type P showed a much larger ability to transform to cristobalite at lower temperatures than type A. Type P had a larger amount of alkali and alkaline earth impurities. This could have enhanced the transformation to cristobalite. For quartz type A the amount of cristobalite was larger at 1600 °C than 1700 °C. This can also be seen for some of the samples of type P at shorter holding times

Phase Transformations from Quartz to Cristobalite

Two quartz types used in the silicon and ferrosilicon industry were heated to temperatures of 1600 and 1700 °C. The parameters varied were the temperature and the holding time at maximum temperature. The amount of quartz, cristobalite and intermediate amorphous phase were measured using XRD and the internal standard method. Type P showed a much larger ability to transform to cristobalite at lower temperatures than type A. Type P had a larger amount of alkali and alkaline earth impurities. This could have enhanced the transformation to cristobalite. For quartz type A the amount of cristobalite was larger at 1600 °C than 1700 °C. This can also be seen for some of the samples of type P at shorter holding times.acceptedVersio

A direct proofreader-clamp interaction stabilizes the Pol III replicase in the polymerization mode

<p>Processive DNA synthesis by the alpha epsilon theta core of the Escherichia coli Pol III replicase requires it to be bound to the beta(2) clamp via a site in the a polymerase subunit. How the epsilon proofreading exonuclease subunit influences DNA synthesis by alpha was not previously understood. In this work, bulk assays of DNA replication were used to uncover a non-proofreading activity of epsilon. Combination of mutagenesis with biophysical studies and single-molecule leading-strand replication assays traced this activity to a novel beta-binding site in e that, in conjunction with the site in a, maintains a closed state of the alpha epsilon theta-beta(2) replicase in the polymerization mode of DNA synthesis. The epsilon-beta interaction, selected during evolution to be weak and thus suited for transient disruption to enable access of alternate polymerases and other clamp binding proteins, therefore makes an important contribution to the network of protein-protein interactions that finely tune stability of the replicase on the DNA template in its various conformational states.</p>

Anatomy of a DNA replication fork revealed by reconstitution of SV40 DNA replication in vitro

Complete enzymatic replication of DNA from the simian virus 40 origin has been reconstituted with T antigen and highly purified cellular proteins. DNA polymerase-alpha/primase functions primarily to synthesize RNA-DNA primers for initiation of DNA replication at the origin and for priming each Okazaki fragment. A polymerase switching mechanism requiring replication factor C and the proliferating cell nuclear antigen allows two molecules of DNA polymerase-delta to replicate both strands of the double helix conjointly