Rmi1 stimulates decatenation of double Holliday junctions during dissolution by Sgs1-Top3

double Holliday junction (dHJ) is a central intermediate of homologous recombination that can be processed to yield crossover or non-crossover recombination products. To preserve genomic integrity, cells possess mechanisms to avoid crossing over. We show that Saccharomyces cerevisiae Sgs1 and Top3 proteins are sufficient to migrate and disentangle a dHJ to produce exclusively non-crossover recombination products, in a reaction termed "dissolution." We show that Rmi1 stimulates dHJ dissolution at low Sgs1-Top3 protein concentrations, although it has no effect on the initial rate of Holliday junction (HJ) migration. Rmi1 serves to stimulate DNA decatenation, removing the last linkages between the repaired and template DNA molecules. Dissolution of a dHJ is a highly efficient and concerted alternative to nucleolytic resolution that prevents crossing over of chromosomes during recombinational DNA repair in mitotic cells and thereby contributes to genomic integrity

Opto-Electronic Characterization of Three Dimensional Topological Insulators

We demonstrate that the terahertz/infrared radiation induced photogalvanic effect, which is sensitive to the surface symmetry and scattering details, can be applied to study the high frequency conductivity of the surface states in (Bi1-xSbx)2Te3 based three dimensional (3D) topological insulators (TI). In particular, measuring the polarization dependence of the photogalvanic current and scanning with a micrometre sized beam spot across the sample, provides access to (i) topographical inhomogeneity's in the electronic properties of the surface states and (ii) the local domain orientation. An important advantage of the proposed method is that it can be applied to study TIs at room temperature and even in materials with a high electron density of bulk carriers.Comment: 6 pages, 4 figure

Room temperature high frequency transport of Dirac fermions in epitaxially grown Sb_2Te_3 based topological insulators

We report on the observation of photogalvanic effects in epitaxially grown Sb_2Te_3 three-dimensional (3D) topological insulators (TI). We show that asymmetric scattering of Dirac electrons driven back and forth by the terahertz electric field results in a dc electric current. Due to the "symmetry filtration" the dc current is generated in the surface electrons only and provides an opto-electronic access to probe the electric transport in TI, surface domains orientation and details of electron scattering even in 3D TI at room temperature where conventional surface electron transport is usually hindered by the high carrier density in the bulk

Characterization of unwanted noise in realistic cavities

The problem of the description of absorption and scattering losses in high-Q cavities is studied. The considerations are based on quantum noise theories, hence the unwanted noise associated with scattering and absorption is taken into account by introduction of additional damping and noise terms in the quantum Langevin equations and input--output relations. Completeness conditions for the description of the cavity models obtained in this way are studied and corresponding replacement schemes are discussed.Comment: Contribution to XI International Conference on Quantum Optics, Minsk, Belarus, 26-31 May, 200

BLM and RMI1 alleviate RPA inhibition of topoIIIα decatenase activity

RPA is a single-stranded DNA binding protein that physically associates with the BLM complex. RPA stimulates BLM helicase activity as well as the double Holliday junction dissolution activity of the BLM-topoisomerase IIIα complex. We investigated the effect of RPA on the ssDNA decatenase activity of topoisomerase IIIα. We found that RPA and other ssDNA binding proteins inhibit decatenation by topoisomerase IIIα. Complex formation between BLM, TopoIIIα, and RMI1 ablates inhibition of decatenation by ssDNA binding proteins. Together, these data indicate that inhibition by RPA does not involve species-specific interactions between RPA and BLM-TopoIIIα-RMI1, which contrasts with RPA modulation of double Holliday junction dissolution. We propose that topoisomerase IIIα and RPA compete to bind to single-stranded regions of catenanes. Interactions with BLM and RMI1 enhance toposiomerase IIIα activity, promoting decatenation in the presence of RPA

Epidermal Growth Factor–PEG Functionalized PAMAM-Pentaethylenehexamine Dendron for Targeted Gene Delivery Produced by Click Chemistry

Aim of this study was the site-specific conjugation of an epidermal growth factor (EGF)-polyethylene glycol (PEG) chain by click chemistry onto a poly(amido amine) (PAMAM) dendron, as a key step toward defined multifunctional carriers for targeted gene delivery. For this purpose, at first propargyl amine cored PAMAM dendrons with ester ends were synthesized. The chain terminal ester groups were then modified by oligoamines with different secondary amino densities. The oligoamine-modified PAMAM dendrons were well biocompatible, as demonstrated in cytotoxicity assays. Among the different oligoamine-modified dendrons, PAMAM-pentaethylenehexamine (PEHA) dendron polyplexes displayed the best gene transfer ability. Conjugation of PAMAM-PEHA dendron with PEG spacer was conducted via click reaction, which was performed before amidation with PEHA. The resultant PEG-PAMAM-PEHA copolymer was then coupled with EGF ligand. pDNA transfections in HuH-7 hepatocellular carcinoma cells showed a 10-fold higher efficiency with the polyplexes containing conjugated EGF as compared to the ligand-free ones, demonstrating the concept of ligand targeting. Overall gene transfer efficiencies, however, were moderate, suggesting that additional measures for overcoming subsequent intracellular bottlenecks in delivery have to be taken

Improved methods for creating migratable Holliday junction substrates

Previously, we published a method for creating a novel DNA substrate, the double Holliday junction substrate. This substrate contains two Holliday junctions that are mobile, topologically constrained and separated by a distance comparable with conversion tract lengths. Although useful for studying late stage homologous recombination in vitro, construction of the substrate requires significant effort. In particular, there are three bottlenecks: (i) production of large quantities of single-stranded DNA; (ii) the loss of a significant portion of the DNA following the recombination step; and (iii) the loss of DNA owing to inefficient gel extraction. To address these limitations, we have made the following changes to the protocol: (i) use of a helper plasmid, rather than exogenous helper phage, to produce single-stranded DNA; (ii) use of the unidirectional ϕC31 integrase system in place of the bidirectional Cre recombinase reaction; and (iii) gel extraction by DNA diffusion. Here, we describe the changes made to the materials and methods and characterize the substrates that can be produced, including migratable single Holliday junctions, hemicatenanes and a quadruple Holliday junction substrate

The AUSTRAL VLBI Observing Program

The AUSTRAL observing program was started in 2011, performing geodetic and astrometric very long baseline interferometry (VLBI) sessions using the new Australian AuScope VLBI antennas at Hobart, Katherine, and Yarragadee, with contribution from the Warkworth (New Zealand) 12 m and Hartebeesthoek (South Africa) 15 m antennas to make a southern hemisphere array of telescopes with similar design and capability. Designed in the style of the next-generation VLBI system, these small and fast antennas allow for a new way of observing, comprising higher data rates and more observations than the standard observing sessions coordinated by the International VLBI Service for Geodesy and Astrometry (IVS). In this contribution, the continuous development of the AUSTRAL sessions is described, leading to an improvement of the results in terms of baseline length repeatabilities by a factor of two since the start of this program. The focus is on the scheduling strategy and increased number of observations, aspects of automated operation, and data logistics, as well as results of the 151 AUSTRAL sessions performed so far. The high number of the AUSTRAL sessions makes them an important contributor to VLBI end-products, such as the terrestrial and celestial reference frames and Earth orientation parameters. We compare AUSTRAL results with other IVS sessions and discuss their suitability for the determination of baselines, station coordinates, source coordinates, and Earth orientation parameters

Darboux polynomials for Lotka-Volterra systems in three dimensions

We consider Lotka-Volterra systems in three dimensions depending on three real parameters. By using elementary algebraic methods we classify the Darboux polynomials (also known as second integrals) for such systems for various values of the parameters, and give the explicit form of the corresponding cofactors. More precisely, we show that a Darboux polynomial of degree greater than one is reducible. In fact, it is a product of linear Darboux polynomials and first integrals.Comment: 16 page