26 research outputs found

    The consequences of replicating in the wrong orientation: Bacterial chromosome duplication without an active replication origin

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    Chromosome replication is regulated in all organisms at the assembly stage of the replication machinery at specific origins. In Escherichia coli the DnaA initiator protein regulates the assembly of replication forks at oriC. This regulation can be undermined by defects in nucleic acid meta¬bolism. In cells lacking RNase HI replication initiates indepen¬dently of DnaA and oriC, presumably at persisting R-loops. A similar mechanism was assumed for origin-independent synthesis in cells lacking RecG. However, recently we suggested that this synthesis initiates at intermediates resulting from replication fork fusions. Here we present data suggesting that in cells lacking RecG or RNase HI origin-independent synthesis arises by different mechanisms, indicative of these two proteins having different roles in vivo. Our data support the idea that RNase HI processes R-loops, while RecG is required to process replication fork fusion intermediates. However, regardless of how origin-independent synthesis is initiated, a fraction of forks will proceed in an orientation opposite to normal. We show that the resulting head-on encounters with transcription threaten cell viability, especially if taking place in highly-transcribed areas. Thus, despite their different functions, RecG and RNase HI are both important factors for maintaining replication control and orientation. Their absence causes severe replication problems, highlighting the advantages of the normal chromosome arrangement, which exploits a single origin to control the number of forks and their orientation relative to transcription, and a defined termination area to contain fork fusions. Any changes to this arrangement endanger cell cycle control, chromosome dynamics and, ultimately, cell viability.This work was supported by the Royal Society (RG110414 to C.J.R.) and The Biotechnology and Biological Sciences Research Council (BB/K015729/1 to C.J.R.)

    Origins left, right and centre: increasing the number of initiation sites in the Escherichia coli chromosome

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    © 2018 by the authors. The bacterium Escherichia coli contains a single circular chromosome with a defined architecture. DNA replication initiates at a single origin called oriC. Two replication forks are assembled and proceed in opposite directions until they fuse in a specialised zone opposite the origin. This termination area is flanked by polar replication fork pause sites that allow forks to enter, but not to leave. Thus, the chromosome is divided into two replichores, each replicated by a single replication fork. Recently, we analysed the replication parameters in E. coli cells, in which an ectopic origin termed oriZ was integrated in the right-hand replichore. Two major obstacles to replication were identified: (1) head-on replication–transcription conflicts at highly transcribed rrn operons, and (2) the replication fork trap. Here, we describe replication parameters in cells with ectopic origins, termed oriX and oriY, integrated into the left-hand replichore, and a triple origin construct with oriX integrated in the left-hand and oriZ in the right-hand replichore. Our data again highlight both replication–transcription conflicts and the replication fork trap as important obstacles to DNA replication, and we describe a number of spontaneous large genomic rearrangements which successfully alleviate some of the problems arising from having an additional origin in an ectopic location. However, our data reveal additional factors that impact efficient chromosome duplication, highlighting the complexity of chromosomal architecture

    Functional traits and phenotypic plasticity modulate species coexistence across contrasting climatic conditions

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    Functional traits are expected to modulate plant competitive dynamics. However, how traits and their plasticity in response to contrasting environments connect with the mechanisms determining species coexistence remains poorly understood. Here, we couple field experiments under two contrasting climatic conditions to a plant population model describing competitive dynamics between 10 annual plant species in order to evaluate how 19 functional traits, covering physiological, morphological and reproductive characteristics, are associated with species’ niche and fitness differences. We find a rich diversity of univariate and multidimensional associations, which highlight the primary role of traits related to water- and lightuse- efficiency for modulating the determinants of competitive outcomes. Importantly, such traits and their plasticity promote species coexistence across climatic conditions by enhancing stabilizing niche differences and by generating competitive trade-offs between species. Our study represents a significant advance showing how leading dimensions of plant function connect to the mechanisms determining the maintenance of biodiversity

    Enhanced thylakoid photoprotection can increase yield and canopy radiation use efficiency in rice

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    High sunlight can raise plant growth rates but can potentially cause cellular damage. The likelihood of deleterious effects is lowered by a sophisticated set of photoprotective mechanisms, one of the most important being the controlled dissipation of energy from chlorophyll within photosystem II (PSII) measured as non-photochemical quenching (NPQ). Although ubiquitous, the role of NPQ in plant productivity remains uncertain because it momentarily reduces the quantum efficiency of photosynthesis. Here we used plants overexpressing the gene encoding a central regulator of NPQ, the protein PsbS, within a major crop species (rice) to assess the effect of photoprotection at the whole canopy scale. We accounted for canopy light interception, to our knowledge for the first time in this context. We show that in comparison to wild-type plants, psbS overexpressors increased canopy radiation use efficiency and grain yield in fluctuating light, demonstrating that photoprotective mechanisms should be altered to improve rice crop productivity

    Toward a General Model for the Evolution of DNA Replication in Three Domains of Life

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    Nothing is more fundamental to life than the ability to reproduce and duplicate the information cells store in their genomes. The mechanism of duplication of DNA has been conserved from prokaryotes to eukaryotes. The aim of the study was to quantify which evolutionary forces could produce the pattern of genome replication architecture observed in present-day organisms. This was achieved using an evolutionary simulation, combining random genome sequence shuffling, mutation, selection and the mathematical modeling of DNA replication. We have found parameter values which explained evolutionary pressures of DNA replication in E.coli, P.calidifontis and S. cerevisae. Surprisingly, the results of the evolutionary simulation suggests that for a fixed cost per replication origin it is more advantageous for genomes to reduce the number of replication origins under increasing uncertainty in origin activation timing

    Nonlinearity in the interaction of ultrasound contrast agents under the periodic acoustic forcing

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    Ultrasound contrast agents, a type of specially coated gas-filled microbubbles, provide a mean for the imaging of vascular volume and flow rate directly and in real time (Beche

    THE ANALYSIS OF CHEMICAL KINETICS EQUATIONS WITH TIME DELAY

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    Statistical methods for linking geostatistical maps and transmission models: Application to lymphatic filariasis in East Africa

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    Infectious diseases remain one of the major causes of human mortality and suffering. Mathematical models have been established as an important tool for capturing the features that drive the spread of the disease, predicting the progression of an epidemic and hence guiding the development of strategies to control it. Another important area of epidemiological interest is the development of geostatistical methods for the analysis of data from spatially referenced prevalence surveys. Maps of prevalence are useful, not only for enabling a more precise disease risk stratification, but also for guiding the planning of more reliable spatial control programmes by identifying affected areas. Despite the methodological advances that have been made in each area independently, efforts to link transmission models and geostatistical maps have been limited. Motivated by this fact, we developed a Bayesian approach that combines fine-scale geostatistical maps of disease prevalence with transmission models to provide quantitative, spatially-explicit projections of the current and future impact of control programs against a disease. These estimates can then be used at a local level to identify the effectiveness of suggested intervention schemes and allow investigation of alternative strategies. The methodology has been applied to lymphatic filariasis in East Africa to provide estimates of the impact of different intervention strategies against the disease

    Integrating geostatistical maps and infectious disease transmission models using Adaptive Multiple Importance Sampling

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    The Adaptive Multiple Importance Sampling algorithm (AMIS) is an iterative technique which recycles samples from all previous iterations in order to improve the efficiency of the proposal distribution. We have formulated a new statistical framework, based on AMIS, to take the output from a geostatistical model of infectious disease prevalence, incidence or relative risk, and project it forward in time under a mathematical model for transmission dynamics. We adapted the AMIS algorithm so that it can sample from multiple targets simultaneously by changing the focus of the adaptation at each iteration. By comparing our approach against the standard AMIS algorithm, we showed that these novel adaptations greatly improve the efficiency of the sampling. We tested the performance of our algorithm on four case studies: ascariasis in Ethiopia, onchocerciasis in Togo, human immunodeficiency virus (HIV) in Botswana, and malaria in the Democratic Republic of the Congo
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