Imaging and energetics of single SSB-ssDNA molecules reveal intramolecular condensation and insight into RecOR function.

Escherichia coli single-stranded DNA (ssDNA) binding protein (SSB) is the defining bacterial member of ssDNA binding proteins essential for DNA maintenance. SSB binds ssDNA with a variable footprint of ∼30-70 nucleotides, reflecting partial or full wrapping of ssDNA around a tetramer of SSB. We directly imaged single molecules of SSB-coated ssDNA using total internal reflection fluorescence (TIRF) microscopy and observed intramolecular condensation of nucleoprotein complexes exceeding expectations based on simple wrapping transitions. We further examined this unexpected property by single-molecule force spectroscopy using magnetic tweezers. In conditions favoring complete wrapping, SSB engages in long-range reversible intramolecular interactions resulting in condensation of the SSB-ssDNA complex. RecO and RecOR, which interact with SSB, further condensed the complex. Our data support the idea that RecOR--and possibly other SSB-interacting proteins-function(s) in part to alter long-range, macroscopic interactions between or throughout nucleoprotein complexes by microscopically altering wrapping and bridging distant sites

Direct imaging of RecA nucleation and growth on single molecules of SSB-coated ssDNA.

Escherichia coli RecA is the defining member of a ubiquitous class of DNA strand-exchange proteins that are essential for homologous recombination, a pathway that maintains genomic integrity by repairing broken DNA. To function, filaments of RecA must nucleate and grow on single-stranded DNA (ssDNA) in direct competition with ssDNA-binding protein (SSB), which rapidly binds and continuously sequesters ssDNA, kinetically blocking RecA assembly. This dynamic self-assembly on a DNA lattice, in competition with another protein, is unique for the RecA family compared to other filament-forming proteins such as actin and tubulin. The complexity of this process has hindered our understanding of RecA filament assembly because ensemble measurements cannot reliably distinguish between the nucleation and growth phases, despite extensive and diverse attempts. Previous single-molecule assays have measured the nucleation and growth of RecA--and its eukaryotic homologue RAD51--on naked double-stranded DNA and ssDNA; however, the template for RecA self-assembly in vivo is SSB-coated ssDNA. Using single-molecule microscopy, here we directly visualize RecA filament assembly on single molecules of SSB-coated ssDNA, simultaneously measuring nucleation and growth. We establish that a dimer of RecA is required for nucleation, followed by growth of the filament through monomer addition, consistent with the finding that nucleation, but not growth, is modulated by nucleotide and magnesium ion cofactors. Filament growth is bidirectional, albeit faster in the 5'→3' direction. Both nucleation and growth are repressed at physiological conditions, highlighting the essential role of recombination mediators in potentiating assembly in vivo. We define a two-step kinetic mechanism in which RecA nucleates on transiently exposed ssDNA during SSB sliding and/or partial dissociation (DNA unwrapping) and then the RecA filament grows. We further demonstrate that the recombination mediator protein pair, RecOR (RecO and RecR), accelerates both RecA nucleation and filament growth, and that the introduction of RecF further stimulates RecA nucleation

Statistical interaction modeling of bovine herd behaviors

While there has been interest in modeling the group behavior of herds or flocks, much of this work has focused on simulating their collective spatial motion patterns which have not accounted for individuality in the herd and instead assume a homogenized role for all members or sub-groups of the herd. Animal behavior experts have noted that domestic animals exhibit behaviors that are indicative of social hierarchy: leader/follower type behaviors are present as well as dominance and subordination, aggression and rank order, and specific social affiliations may also exist. Both wild and domestic cattle are social species, and group behaviors are likely to be influenced by the expression of specific social interactions. In this paper, Global Positioning System coordinate fixes gathered from a herd of beef cows tracked in open fields over several days at a time are utilized to learn a model that focuses on the interactions within the herd as well as its overall movement. Using these data in this way explores the validity of existing group behavior models against actual herding behaviors. Domain knowledge, location geography and human observations, are utilized to explain the causes of these deviations from this idealized behavior

Difficulty of distinguishing product states locally

Non-locality without entanglement is a rather counter-intuitive phenomenon in which information may be encoded entirely in product (unentangled) states of composite quantum systems in such a way that local measurement of the subsystems is not enough for optimal decoding. For simple examples of pure product states, the gap in performance is known to be rather small when arbitrary local strategies are allowed. Here we restrict to local strategies readily achievable with current technology; those requiring neither a quantum memory nor joint operations. We show that, even for measurements on pure product states there can be a large gap between such strategies and theoretically optimal performance. Thus even in the absence of entanglement physically realizable local strategies can be far from optimal for extracting quantum information.Comment: 5 pages, 1 figur

Strategic Operations Research and the Edelman Prize Finalist Applications 1989-1998

In an earlier study we examined the available evidence on the Edelman Prize finalist applications 1989-1998. This study concluded that 13 of the 42 private sector applications provide examples of strategic operations research (SOR) when SOR is defined as operations research that creates a sustainable competitive advantage. In a follow-up study we tested our classifications, gathering longitudinal information on the continued success of the Edelman applications. We contacted people who were familiar with all the private sector applications that were Edelman finalists 1989-1996 and had at least five years of history since the competition. We describe the post-Edelman history of these applications and use this data to reassess their strategic role. We found that the longitudinal data provides evidence to support our original classification, but also suggests that several additional applications were more strategic than was originally apparent. We conclude that almost 60% (20 of 34) of these applications created a sustainable competitive advantage for their firms and provide examples of SOR

Generating nonclassical correlations without fully aligning measurements

We investigate the scenario where spatially separated parties perform measurements in randomly chosen bases on an N-partite Greenberger-Horne-Zeilinger state. We show that without any alignment of the measurements, the observers will obtain correlations that violate a Bell inequality with a probability that rapidly approaches 1 as N increases and that this probability is robust against noise. We also prove that restricting these randomly chosen measurements to a plane perpendicular to a common direction will always generate correlations that violate some Bell inequality. Specifically, if each observer chooses their two measurements to be locally orthogonal, then the N observers will violate one of two Bell inequalities by an amount that increases exponentially with N. These results are also robust against noise and perturbations of each observer's reference direction from the common direction.Comment: v2: Essentially published version (with typos fixed, results updated in Table 2 and Figure 4 replaced); v1: 16 pages, 5 figures, 2 tables, comments welcom

Differential regulation of insulin-like growth factor binding protein-1 and -2 by insulin in the baboon (Papio anubis) endometrium

<p>Abstract</p> <p>Background</p> <p>The purpose of this study was to examine the effect of insulin on expression and synthesis of IGFBP-1 and IGFBP-2 in the baboon endometrium in vitro.</p> <p>Methods</p> <p>Baboon endometrial explants collected from cycling, ovariectomized, steroid-treated, simulated-pregnant and pregnant animals were cultured for 48 h in the presence or absence of insulin, with or without estradiol, progesterone and hCG.</p> <p>Results</p> <p>Insulin clearly inhibited IGFBP-1 production and mRNA expression in a time- and dose-dependent manner, whereas IGFBP-2 synthesis was not significantly affected. The inhibitory effects of insulin on IGFBP-1 were more evident in explants of non-pregnant tissue or tissue away from the implantation site. In the absence of insulin, synthesis of IGFBP-1 was induced in explants with low levels of de novo synthesis whereas IGFBP-2 synthesis was inhibited. This effect was potentiated by steroids and hCG in the explant cultures.</p> <p>Conclusion</p> <p>Insulin differentially regulates endometrial IGFBP-1 and IGFBP-2 secretion in the baboon.</p

Comparative and Evolutionary Analysis of the Bacterial Homologous Recombination Systems

Homologous recombination is a housekeeping process involved in the maintenance of chromosome integrity and generation of genetic variability. Although detailed biochemical studies have described the mechanism of action of its components in model organisms, there is no recent extensive assessment of this knowledge, using comparative genomics and taking advantage of available experimental data on recombination. Using comparative genomics, we assessed the diversity of recombination processes among bacteria, and simulations suggest that we missed very few homologs. The work included the identification of orthologs and the analysis of their evolutionary history and genomic context. Some genes, for proteins such as RecA, the resolvases, and RecR, were found to be nearly ubiquitous, suggesting that the large majority of bacterial genomes are capable of homologous recombination. Yet many genomes show incomplete sets of presynaptic systems, with RecFOR being more frequent than RecBCD/AddAB. There is a significant pattern of co-occurrence between these systems and antirecombinant proteins such as the ones of mismatch repair and SbcB, but no significant association with nonhomologous end joining, which seems rare in bacteria. Surprisingly, a large number of genomes in which homologous recombination has been reported lack many of the enzymes involved in the presynaptic systems. The lack of obvious correlation between the presence of characterized presynaptic genes and experimental data on the frequency of recombination suggests the existence of still-unknown presynaptic mechanisms in bacteria. It also indicates that, at the moment, the assessment of the intrinsic stability or recombination isolation of bacteria in most cases cannot be inferred from the identification of known recombination proteins in the genomes