Rad51/Dmc1 paralogs and mediators oppose DNA helicases to limit hybrid DNA formation and promote crossovers during meiotic recombination

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. ACKNOWLEDGMENTS We are grateful to J ¨urg Kohli, Ramsay J. McFarlane, Paul Russell, Gerald R. Smith, Walter W. Steiner and the National BioResource Project (NBRP) Japan for providing strains and to C. Bryer for technical assistance. FUNDING Wellcome Trust [090767/Z/09/Z to M.C.W.]; College of Life Sciences and Medicine, University of Aberdeen [to A.L., in part]. Funding for open access charge: Wellcome TrustPeer reviewedPublisher PD

A failure of meiotic chromosome segregation in a fbh1Δ mutant correlates with persistent Rad51-DNA associations

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MHF1–2/CENP-S-X performs distinct roles in centromere metabolism and genetic recombination

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Biofilm and planktonic bacterial and fungal communities transforming high molecular weight polycyclic aromatic hydrocarbons.

High molecular weight polycyclic aromatic hydrocarbons (HMW-PAHs) are natural components of fossil fuels that are carcinogenic and persistent in the environment, particularly in oil sands process-affected water (OSPW). Their hydrophobicity and tendency to adsorb to organic matter result in low bioavailability and high recalcitrance to degradation. Despite the importance of microbes for environmental remediation, little is known about those involved in HMW-PAH transformations. Here, we investigated the transformation of HMW-PAHs using samples of OSPW, and compared the bacterial and fungal community composition attached to hydrophobic filters and in suspension. It was anticipated that the hydrophobic filters with sorbed HMW-PAHs would select for microbes that specialise in adhesion. Over 33 days more pyrene was removed (75% ± 11.7) than the five-ring PAHs benzo[a]pyrene (44% ± 13.6) and benzo[b]fluoranthene (41% ± 12.6). For both bacteria and fungi, the addition of PAHs led to a shift in community composition, but thereafter the major factor determining the fungal community composition was whether they were in the planktonic phase or attached to filters. In contrast, the major determinant of the bacterial community composition was the nature of the PAH serving as the carbon source. The main bacteria enriched by HMW-PAHs were Pseudomonas, Bacillus and Microbacterium species. This report demonstrates that OSPW harbour microbial communities with the capacity to transform HMW-PAHs. Furthermore, the provision of suitable surfaces that encourage PAH sorption and microbial adhesion select for different fungal and bacterial species with the potential for HMW-PAH degradation

Efficient Second Strand Cleavage during Holliday Junction Resolution by RuvC Requires Both Increased Junction Flexibility and an Exposed 5′ Phosphate

BACKGROUND: Holliday junction (HJ) resolution is a critical step during homologous recombination. In Escherichia coli this job is performed by a member of the RNase H/Integrase superfamily called RuvC, whereas in Schizosaccharomyces pombe it has been attributed to the XPF family member Mus81-Eme1. HJ resolution is achieved through the sequential cleavage of two strands of like polarity at or close to the junction crossover point. RuvC functions as a dimer, whereas Mus81-Eme1 is thought to function as a dimer of heterodimers. However, in both cases the multimer contains two catalytic sites, which act independently and sequentially during the resolution reaction. To ensure that both strands are cleaved before the nuclease dissociates from the junction, the rate of second strand cleavage is greatly enhanced compared to that of the first. The enhancement of second strand cleavage has been attributed to the increased flexibility of the nicked HJ, which would facilitate rapid engagement of the second active site and scissile bond. Here we have investigated whether other properties of the nicked HJ are important for enhancing second strand cleavage. PRINCIPAL FINDINGS: A comparison of the efficiency of cleavage of nicked HJs with and without a 5' phosphate at the nick site shows that a 5' phosphate is required for most of the enhancement of second strand cleavage by RuvC. In contrast Mus81-Eme1 cleaves nicked HJs with and without a 5' phosphate with equal efficiency, albeit there are differences in cleavage site selection. CONCLUSIONS: Our data show that efficient HJ resolution by RuvC depends on the 5' phosphate revealed by incision of the first strand. This is a hitherto unappreciated factor in promoting accelerated second strand cleavage. However, a 5' phosphate is not a universal requirement since efficient cleavage by Mus81-Eme1 appears to depend solely on the increased junction flexibility that is developed by the first incision

The RecQ DNA helicase Rqh1 constrains Exonuclease 1-dependent recombination at stalled replication forks

Acknowledgements We thank Benoit Arcangioli, Andrei Kuzminov and Shao-Win Wang for providing strains/plasmids. This work was supported by grants 057586/Z/99/A and 090767/Z/09/Z from the Wellcome Trust.Peer reviewedPublisher PD

The human Holliday junction resolvase GEN1 rescues the meiotic phenotype of a Schizosaccharomyces pombe mus81 mutant

A key step in meiotic recombination involves the nucleolytic resolution of Holliday junctions to generate crossovers. Although the enzyme that performs this function in human cells is presently unknown, recent studies led to the identification of the XPG-family endonuclease GEN1 that promotes Holliday junction resolution in vitro, suggesting that it may perform a related function in vivo. Here, we show that ectopic expression of GEN1 in fission yeast mus81Δ strains results in Holliday junction resolution and crossover formation during meiosis

A Learning Automata Based Solution to Service Selection in Stochastic Environments

With the abundance of services available in today’s world, identifying those of high quality is becoming increasingly difficult. Reputation systems can offer generic recommendations by aggregating user provided opinions about service quality, however, are prone to ballot stuffing and badmouthing . In general, unfair ratings may degrade the trustworthiness of reputation systems, and changes in service quality over time render previous ratings unreliable. In this paper, we provide a novel solution to the above problems based on Learning Automata (LA), which can learn the optimal action when operating in unknown stochastic environments. Furthermore, they combine rapid and accurate convergence with low computational complexity. In additional to its computational simplicity, unlike most reported approaches, our scheme does not require prior knowledge of the degree of any of the above mentioned problems with reputation systems. Instead, it gradually learns which users provide fair ratings, and which users provide unfair ratings, even when users unintentionally make mistakes. Comprehensive empirical results show that our LA based scheme efficiently handles any degree of unfair ratings (as long as ratings are binary). Furthermore, if the quality of services and/or the trustworthiness of users change, our scheme is able to robustly track such changes over time. Finally, the scheme is ideal for decentralized processing. Accordingly, we believe that our LA based scheme forms a promising basis for improving the performance of reputation systems in general