N-body Models of Rotating Globular Clusters

We have studied the dynamical evolution of rotating globular clusters with direct $N$-body models. Our initial models are rotating King models; we obtained results for both equal-mass systems and systems composed out of two mass components. Previous investigations using a Fokker-Planck solver have revealed that rotation has a noticeable influence on stellar systems like globular clusters, which evolve by two-body relaxation. In particular, it accelerates their dynamical evolution through the gravogyro instability. We have validated the occurence of the gravogyro instability with direct $N$-body models. In the case of systems composed out of two mass components, mass segregation takes place, which competes with the rotation in the acceleration of the core collapse. The "accelerating" effect of rotation has not been detected in our isolated two-mass $N$-body models. Last, but not least, we have looked at rotating $N$-body models in a tidal field within the tidal approximation. It turns out that rotation increases the escape rate significantly. A difference between retrograde and prograde rotating star clusters occurs with respect to the orbit of the star cluster around the Galaxy, which is due to the presence of a ``third integral'' and chaotic scattering, respectively.Comment: 16 pages, 17 figures, accepted by MNRA

Radial velocities in the globular cluster omega Centauri

We have used the ARGUS multi-object spectrometer at the CTIO 4m Blanco telescope to obtain 2756 radial velocity measurements for 1966 individual stars in the globular cluster omega Centauri brighter than blue photographic magnitude of about 16.5. Of these, 1589 stars are cluster members. A comparison with two independent radial velocity studies, carried out by Suntzeff & Kraft and by Mayor et al., demonstrates that the median error of our measurements is below 2 km/s for the stars brighter than B-magnitude 15, which constitute the bulk of the sample. The observed velocity dispersion decreases from about 15 km/s in the inner few arcmin to about 6 km/s at a radius of 25 arcmin. The cluster shows significant rotation, with a maximum amplitude of about 6 km/s in the radial zone between 6 and 10 arcmin. In a companion paper by van de Ven et al., we correct these radial velocities for the perspective rotation caused by the space motion of the cluster, and combine them with the internal proper motions of nearly 8000 cluster members measured by van Leeuwen et al., to construct a detailed dynamical model of omega Centauri and to measure its distance.Comment: 10 pages (7 figures), accepted for publication in A&

RNase H2, mutated in Aicardi-Goutières syndrome, promotes LINE-1 retrotransposition

Long INterspersed Element class 1 (LINE-1) elements are a type of abundant retrotransposons active in mammalian genomes. An average human genome contains ~100 retrotransposition-competent LINE-1s, whose activity is influenced by the combined action of cellular repressors and activators. TREX1, SAMHD1 and ADAR1 are known LINE-1 repressors and when mutated cause the autoinflammatory disorder Aicardi-Goutières syndrome (AGS). Mutations in RNase H2 are the most common cause of AGS, and its activity was proposed to similarly control LINE-1 retrotransposition. It has therefore been suggested that increased LINE-1 activity may be the cause of aberrant innate immune activation in AGS. Here, we establish that, contrary to expectations, RNase H2 is required for efficient LINE-1 retrotransposition. As RNase H1 overexpression partially rescues the defect in RNase H2 null cells, we propose a model in which RNase H2 degrades the LINE-1 RNA after reverse transcription, allowing retrotransposition to be completed. This also explains how LINE-1 elements can retrotranspose efficiently without their own RNase H activity. Our findings appear to be at odds with LINE-1-derived nucleic acids driving autoinflammation in AGS.M.B.-G. is funded by a “Formacion Profesorado Universitario” (FPU) PhD fellowship from the Government of Spain (MINECO, Ref FPU15/03294), and this paper is part of her thesis project (“Epigenetic control of the mobility of a human retrotransposon”). R.V.-A. is funded by a PFIS Fellowship from the Government of Spain (ISCiii, FI16/00413). O.M. is funded by an EMBO Long-Term Fellowship (ALTF 7-2015), the European Commission FP7 (Marie Curie Actions, LTFCOFUND2013, GA-2013-609409) and the Swiss National Science Foundation (P2ZHP3_158709). S.R.H. is funded by the Government of Spain (MINECO, RYC-2016-21395 and SAF2015-71589-P). A.P.J’s laboratory is supported by the UK Medical Research Council (MRC University Unit grant U127527202). J.L.G.P’s laboratory is supported by CICEFEDER- P12-CTS-2256, Plan Nacional de I+D+I 2008-2011 and 2013-2016 (FISFEDER- PI14/02152), PCIN-2014-115-ERA-NET NEURON II, the European Research Council (ERC-Consolidator ERC-STG-2012-233764), by an International Early Career Scientist grant from the Howard Hughes Medical Institute (IECS-55007420), by The Wellcome Trust-University of Edinburgh Institutional Strategic Support Fund (ISFF2) and by a private donation from Ms Francisca Serrano (Trading y Bolsa para Torpes, Granada, Spain)

When TADs go bad: chromatin structure and nuclear organisation in human disease

Chromatin in the interphase nucleus is organised as a hierarchical series of structural domains, including self-interacting domains called topologically associating domains (TADs). This arrangement is thought to bring enhancers into closer physical proximity with their target genes, which often are located hundreds of kilobases away in linear genomic distance. TADs are demarcated by boundary regions bound by architectural proteins, such as CTCF and cohesin, although much remains to be discovered about the structure and function of these domains. Recent studies of TAD boundaries disrupted in engineered mouse models show that boundary mutations can recapitulate human developmental disorders as a result of aberrant promoter-enhancer interactions in the affected TADs. Similar boundary disruptions in certain cancers can result in oncogene overexpression, and CTCF binding sites at boundaries appear to be hyper-mutated across cancers. Further insights into chromatin organisation, in parallel with accumulating whole genome sequence data for disease cohorts, are likely to yield additional valuable insights into the roles of noncoding sequence variation in human disease

The non-peculiar velocity dispersion profile of the stellar system omega Centauri

We present the results of a survey of radial velocities over a wide region extending from r~10 arcmin out to r~80 arcmin (~1.5 tidal radii) within the massive star cluster omega Centauri. The survey was performed with FLAMES@VLT, to study the velocity dispersion profile in the outer regions of this stellar system. We derived accurate radial velocities for a sample of 2557 newly observed stars, identifying 318 bona-fide cluster red giants. Merging our data with those provided by Pancino et al. (2007), we assembled a final homogeneous sample of 946 cluster members that allowed us to trace the velocity dispersion profile from the center out to r~32 arcmin. The velocity dispersion appears to decrease monotonically over this range, from a central value of sigma_{v}~17.2 Km/s down to a minimum value of sigma_{v}~5.2 Km/s. The observed surface brightness profile, rotation curve, velocity dispersion profile and ellipticity profile are simultaneously well reproduced by a simple dynamical model in which mass follows light, within the classical Newtonian theory of gravitation. The comparison with an N-body model of the evolution of a system mimicking omega Cen during the last 10 orbits into the Galactic potential suggests that (a) the rotation of stars lying in the inner ~20 arcmin of the clusters is not due to the effects of the tidal field of the Milky Way, as hypothized by other authors, and (b) the overall observational scenario is still compatible with the possibility that the outer regions of the cluster are subject to some tidal stirring.Comment: 12 pages, 12 figures, accepted for publication by MNRA

PCNA directs type 2 RNase H activity on DNA replication and repair substrates

Ribonuclease H2 is the major nuclear enzyme degrading cellular RNA/DNA hybrids in eukaryotes and the sole nuclease known to be able to hydrolyze ribonucleotides misincorporated during genomic replication. Mutation in RNASEH2 causes Aicardi–Goutières syndrome, an auto-inflammatory disorder that may arise from nucleic acid byproducts generated during DNA replication. Here, we report the crystal structures of Archaeoglobus fulgidus RNase HII in complex with PCNA, and human PCNA bound to a C-terminal peptide of RNASEH2B. In the archaeal structure, three binding modes are observed as the enzyme rotates about a flexible hinge while anchored to PCNA by its PIP-box motif. PCNA binding promotes RNase HII activity in a hinge-dependent manner. It enhances both cleavage of ribonucleotides misincorporated in DNA duplexes, and the comprehensive hydrolysis of RNA primers formed during Okazaki fragment maturation. In addition, PCNA imposes strand specificity on enzyme function, and by localizing RNase H2 and not RNase H1 to nuclear replication foci in vivo it ensures that RNase H2 is the dominant RNase H activity during nuclear replication. Our findings provide insights into how type 2 RNase H activity is directed during genome replication and repair, and suggest a mechanism by which RNase H2 may suppress generation of immunostimulatory nucleic acids

Chromatin loop anchors are associated with genome instability in cancer and recombination hotspots in the germline

Abstract Background Chromatin loops form a basic unit of interphase nuclear organization, with chromatin loop anchor points providing contacts between regulatory regions and promoters. However, the mutational landscape at these anchor points remains under-studied. Here, we describe the unusual patterns of somatic mutations and germline variation associated with loop anchor points and explore the underlying features influencing these patterns. Results Analyses of whole genome sequencing datasets reveal that anchor points are strongly depleted for single nucleotide variants (SNVs) in tumours. Despite low SNV rates in their genomic neighbourhood, anchor points emerge as sites of evolutionary innovation, showing enrichment for structural variant (SV) breakpoints and a peak of SNVs at focal CTCF sites within the anchor points. Both CTCF-bound and non-CTCF anchor points harbour an excess of SV breakpoints in multiple tumour types and are prone to double-strand breaks in cell lines. Common fragile sites, which are hotspots for genome instability, also show elevated numbers of intersecting loop anchor points. Recurrently disrupted anchor points are enriched for genes with functions in cell cycle transitions and regions associated with predisposition to cancer. We also discover a novel class of CTCF-bound anchor points which overlap meiotic recombination hotspots and are enriched for the core PRDM9 binding motif, suggesting that the anchor points have been foci for diversity generated during recent human evolution. Conclusions We suggest that the unusual chromatin environment at loop anchor points underlies the elevated rates of variation observed, marking them as sites of regulatory importance but also genomic fragility

Polymerase δ replicates both strands after homologous recombination-dependent fork restart

To maintain genetic stability DNA must be replicated only once and replication completed even when individual replication forks are inactivated. Because fork inactivation is common, the passive convergence of an adjacent fork is insufficient to rescue all inactive forks. Thus, eukaryotic cells have evolved homologous recombination-dependent mechanisms to restart persistent inactive forks. Completing DNA synthesis via Homologous Recombination Restarted Replication (HoRReR) ensures cell survival, but at a cost. One such cost is increased mutagenesis caused by HoRReR being more error prone than canonical replication. This increased error rate implies that the HoRReR mechanism is distinct from that of a canonical fork. Here we exploit the fission yeast Schizosaccharomyces pombe to demonstrate that a DNA sequence duplicated by HoRReR during S phase is replicated semi-conservatively, but that both the leading and lagging strands are synthesised by DNA polymerase delta

Ribonuclease H2 mutations induce a cGAS/STING-dependent innate immune response

Aicardi–Goutières syndrome (AGS) provides a monogenic model of nucleic acid‐mediated inflammation relevant to the pathogenesis of systemic autoimmunity. Mutations that impair ribonuclease (RNase) H2 enzyme function are the most frequent cause of this autoinflammatory disorder of childhood and are also associated with systemic lupus erythematosus. Reduced processing of either RNA:DNA hybrid or genome‐embedded ribonucleotide substrates is thought to lead to activation of a yet undefined nucleic acid‐sensing pathway. Here, we establish Rnaseh2b (A174T/A174T) knock‐in mice as a subclinical model of disease, identifying significant interferon‐stimulated gene (ISG) transcript upregulation that recapitulates the ISG signature seen in AGS patients. The inflammatory response is dependent on the nucleic acid sensor cyclic GMP‐AMP synthase (cGAS) and its adaptor STING and is associated with reduced cellular ribonucleotide excision repair activity and increased DNA damage. This suggests that cGAS/STING is a key nucleic acid‐sensing pathway relevant to AGS, providing additional insight into disease pathogenesis relevant to the development of therapeutics for this childhood‐onset interferonopathy and adult systemic autoimmune disorders