Interplay of Mre11 Nuclease with Dna2 plus Sgs1 in Rad51-Dependent Recombinational Repair

The Mre11/Rad50/Xrs2 complex initiates IR repair by binding to the end of a double-strand break, resulting in 5′ to 3′ exonuclease degradation creating a single-stranded 3′ overhang competent for strand invasion into the unbroken chromosome. The nuclease(s) involved are not well understood. Mre11 encodes a nuclease, but it has 3′ to 5′, rather than 5′ to 3′ activity. Furthermore, mutations that inactivate only the nuclease activity of Mre11 but not its other repair functions, mre11-D56N and mre11-H125N, are resistant to IR. This suggests that another nuclease can catalyze 5′ to 3′ degradation. One candidate nuclease that has not been tested to date because it is encoded by an essential gene is the Dna2 helicase/nuclease. We recently reported the ability to suppress the lethality of a dna2Δ with a pif1Δ. The dna2Δ pif1Δ mutant is IR-resistant. We have determined that dna2Δ pif1Δ mre11-D56N and dna2Δ pif1Δ mre11-H125N strains are equally as sensitive to IR as mre11Δ strains, suggesting that in the absence of Dna2, Mre11 nuclease carries out repair. The dna2Δ pif1Δ mre11-D56N triple mutant is complemented by plasmids expressing Mre11, Dna2 or dna2K1080E, a mutant with defective helicase and functional nuclease, demonstrating that the nuclease of Dna2 compensates for the absence of Mre11 nuclease in IR repair, presumably in 5′ to 3′ degradation at DSB ends. We further show that sgs1Δ mre11-H125N, but not sgs1Δ, is very sensitive to IR, implicating the Sgs1 helicase in the Dna2-mediated pathway

Coordination of Nucleases and Helicases during DNA Replication and Double-strand Break Repair

Nucleases and helicases are involved in numerous steps in DNA replication and repair. Nucleases act on intermediates in DNA replication created by DNA polymerases (Chapter 4) and helicases (Chapter 3). They can create substrates for repair as in Okazaki fragment processing (OFP) and homologous recombination. They can also create substrates for activation of a checkpoint response, or participate in downregulation of checkpoints. In the special case of telomere replication, they are also involved in essential processing steps (Chapter 8). Nucleases known to act during DNA replication include Dna2, Rad27, Mre11, Sae2, Exo1, RNaseH, Yen1 andMus81/Mms4. Of these, Dna2, Exo1 and Mre11 are of particular interest because they have been identified as crucial activities that initiate repair of double-strand breaks (DSBs) by homologous recombination and thus form an intrinsic link between DNA replication and repair of DSBs derived from replication fork failure. The action of the nucleases is coordinated with those of a number of helicases and is discussed here in the context of a network of their interactions that combine to maintain genome integrity during DNA replication

Coordination of Nucleases and Helicases during DNA Replication and Double-strand Break Repair

Nucleases and helicases are involved in numerous steps in DNA replication and repair. Nucleases act on intermediates in DNA replication created by DNA polymerases (Chapter 4) and helicases (Chapter 3). They can create substrates for repair as in Okazaki fragment processing (OFP) and homologous recombination. They can also create substrates for activation of a checkpoint response, or participate in downregulation of checkpoints. In the special case of telomere replication, they are also involved in essential processing steps (Chapter 8). Nucleases known to act during DNA replication include Dna2, Rad27, Mre11, Sae2, Exo1, RNaseH, Yen1 andMus81/Mms4. Of these, Dna2, Exo1 and Mre11 are of particular interest because they have been identified as crucial activities that initiate repair of double-strand breaks (DSBs) by homologous recombination and thus form an intrinsic link between DNA replication and repair of DSBs derived from replication fork failure. The action of the nucleases is coordinated with those of a number of helicases and is discussed here in the context of a network of their interactions that combine to maintain genome integrity during DNA replication

Socioeconomic inequalities in outcome of pregnancy and neonatal mortality associated with congenital anomalies: population based study

Objectives To investigate socioeconomic inequalities in outcome of pregnancy and neonatal mortality associated with congenital anomalies

Inviability of a DNA2 deletion mutant is due to the DNA damage checkpoint

Dna2 is a dual polarity exo/endonuclease, and 5' to 3' DNA helicase involved in Okazaki Fragment Processing (OFP) and Double-Strand Break (DSB) Repair. In yeast, DNA2 is an essential gene, as expected for a DNA replication protein. Suppression of the lethality of dna2Δ mutants has been found to occur by two mechanisms: overexpression of RAD27^(scFEN1), encoding a 5' to 3' exo/endo nuclease that processes Okazaki fragments (OFs) for ligation, or deletion of PIF1, a 5' to 3' helicase involved in mitochondrial recombination, telomerase inhibition and OFP. Mapping of a novel, spontaneously arising suppressor of dna2Δ now reveals that mutation of rad9 and double mutation of rad9 mrc1 can also suppress the lethality of dna2Δ mutants. Interaction of dna2Δ and DNA damage checkpoint mutations provides insight as to why dna2Δ is lethal but rad27Δ is not, even though evidence shows that Rad27^(ScFEN1) processes most of the Okazaki fragments, while Dna2 processes only a subset

DNA end resection by Dna2–Sgs1–RPA and its stimulation by Top3–Rmi1 and Mre11–Rad50–Xrs2

The repair of DNA double-strand breaks (DSBs) by homologous recombination requires processing of broken ends. For repair to start, the DSB must first be resected to generate a 3′-single-stranded DNA (ssDNA) overhang, which becomes a substrate for the DNA strand exchange protein, Rad51 (ref. 1). Genetic studies have implicated a multitude of proteins in the process, including helicases, nucleases and topoisomerases. Here we biochemically reconstitute elements of the resection process and reveal that it requires the nuclease Dna2, the RecQ-family helicase Sgs1 and the ssDNA-binding protein replication protein-A (RPA). We establish that Dna2, Sgs1 and RPA constitute a minimal protein complex capable of DNA resection in vitro. Sgs1 helicase unwinds the DNA to produce an intermediate that is digested by Dna2, and RPA stimulates DNA unwinding by Sgs1 in a species-specific manner. Interestingly, RPA is also required both to direct Dna2 nucleolytic activity to the 5′-terminated strand of the DNA break and to inhibit 3′ to 5′ degradation by Dna2, actions that generate and protect the 3′-ssDNA overhang, respectively. In addition to this core machinery, we establish that both the topoisomerase 3 (Top3) and Rmi1 complex and the Mre11–Rad50–Xrs2 complex (MRX) have important roles as stimulatory components. Stimulation of end resection by the Top3–Rmi1 heterodimer and the MRX proteins is by complex formation with Sgs1 (refs 5, 6), which unexpectedly stimulates DNA unwinding. We suggest that Top3–Rmi1 and MRX are important for recruitment of the Sgs1–Dna2 complex to DSBs. Our experiments provide a mechanistic framework for understanding the initial steps of recombinational DNA repair in eukaryotes

The CSIRO Mk3L climate system model version 1.0 – Part 1: Description and evaluation

The CSIRO Mk3L climate system model is a coupled general circulation model, designed primarily for millennial-scale climate simulations and palaeoclimate research. Mk3L includes components which describe the atmosphere, ocean, sea ice and land surface, and combines computational efficiency with a stable and realistic control climatology. This paper describes the model physics and software, analyses the control climatology, and evaluates the ability of the model to simulate the modern climate. <br><br> Mk3L incorporates a spectral atmospheric general circulation model, a <i>z</i>-coordinate ocean general circulation model, a dynamic-thermodynamic sea ice model and a land surface scheme with static vegetation. The source code is highly portable, and has no dependence upon proprietary software. The model distribution is freely available to the research community. A 1000-yr climate simulation can be completed in around one-and-a-half months on a typical desktop computer, with greater throughput being possible on high-performance computing facilities. <br><br> Mk3L produces realistic simulations of the larger-scale features of the modern climate, although with some biases on the regional scale. The model also produces reasonable representations of the leading modes of internal climate variability in both the tropics and extratropics. The control state of the model exhibits a high degree of stability, with only a weak cooling trend on millennial timescales. Ongoing development work aims to improve the model climatology and transform Mk3L into a comprehensive earth system model

Floating stones off El Hierro, Canary Islands: xenoliths of pre-island sedimentary origin in the early products of the October 2011 eruption

The eruption that started off the south coast of El Hierro, Canary Islands, in October 2011 has emitted intriguing eruption products found floating in the sea. These specimens appeared as floating volcanic "bombs" that have in the meantime been termed "restingolites" (after the close-by village of La Restinga) and exhibit cores of white and porous pumice-like material. Currently the nature and origin of these "floating stones" is vigorously debated among researchers, with important implications for the interpretation of the hazard potential of the ongoing eruption. The "restingolites" have been proposed to be either (i) juvenile high-silica magma (e.g. rhyolite), (ii) remelted magmatic material (trachyte), (iii) altered volcanic rock, or (iv) reheated hyaloclastites or zeolite from the submarine slopes of El Hierro. Here, we provide evidence that supports yet a different conclusion. We have collected and analysed the structure and composition of samples and compared the results to previous work on similar rocks found in the archipelago. Based on their high silica content, the lack of igneous trace element signatures, and the presence of remnant quartz crystals, jasper fragments and carbonate relicts, we conclude that "restingolites" are in fact xenoliths from pre-island sedimentary rocks that were picked up and heated by the ascending magma causing them to partially melt and vesiculate. They hence represent messengers from depth that help us to understand the interaction between ascending magma and crustal lithologies in the Canary Islands as well as in similar Atlantic islands that rest on sediment/covered ocean crust (e.g. Cape Verdes, Azores). The occurrence of these "restingolites" does therefore not indicate the presence of an explosive high-silica magma that is involved in the ongoing eruption

Addressing Profound Disadvantages to Improve Indigenous Health and Reduce Hospitalisation: A Collaborative Community Program in Remote Northern Territory

BACKGROUND: Aboriginal people in rural and remote areas of the Northern Territory of Australia have suffered longstanding issues of homelessness and profound health and social inequities. The town and region of Katherine are particularly impacted by such inequities and have the highest rates of homelessness in Australia, composed almost entirely of Aboriginal people who represent 51% of the total population of 24,000 people. The region is serviced by a 60-bed hospital, and a small cohort of frequent attenders (FAs) represent 11% of the Emergency Department (ED) case load. The vast majority of FAs are Aboriginal and have very high burdens of social inequity and homelessness. FAs are a challenge to efficient and effective use of resources for most hospitals around the world, and investment in programs to address underlying social and chronic health issues contributing to frequent attendance have been demonstrated to be effective. METHODS: These are the interim findings of a prospective cohort study using five sources of linked health and related data to evaluate a community-based case management pilot in a culturally competent framework to support frequent attenders to the Katherine Hospital ED. FAs were defined as people with six or more presentations in 12 preceding months. The intervention composed of a community-based case management program with a multi-agency service delivery addressing underlying vulnerabilities contributing to ED presentations. RESULTS: Among this predominantly Aboriginal cohort (91%), there were high rates of homelessness (64%), food insecurity (60%) and alcohol misuse (64%), limited access to transport, and complex comorbidities (average of 2.8 chronic conditions per client). Following intervention, there was a statistically significant reduction in ED presentations (IRR 0.77, 95% CI 0.69-0.85), increased engagement with primary health care (IRR 1.90, 95% CI 1.78-2.03), and ambulance utilisation (IRR 1.21, 95% CI 1.07-1.38). Reductions in hospital admissions (IRR 0.93, 95% CI 0.77-1.10) and aeromedical retrievals (IRR 0.67, 95% CI 0.35-1.20) were not statistically significant. CONCLUSIONS: This study demonstrates the short-term impacts of community-led case management extending beyond the hospital setting, to address causes of recurrent ED presentations among people with complex social and medical backgrounds. Improving engagement with primary care is a particularly important outcome given the national impetus to reduce preventable hospital admissions