Chandra detection of diffuse X-ray emission from the globular cluster Terzan 5

Terzan 5, a globular cluster (GC) prominent in mass and population of compact objects, is searched for diffuse X-ray emission, as proposed by several models. We analyzed the data of an archival Chandra observation of Terzan 5 to search for extended diffuse X-ray emission outside the half-mass radius of the GC. We removed detected point sources from the data to extract spectra from diffuse regions around Terzan 5. The Galactic background emission was modeled by a 2-temperature thermal component, which is typical for Galactic diffuse emission. We detected significant diffuse excess emission above the particle background level from the whole field-of-view. The surface brightness appears to be peaked at the GC center and decreases smoothly outwards. After the subtraction of particle and Galactic background, the excess spectrum of the diffuse emission between the half-mass radius and 3' can be described by a power-law model with photon index $\Gamma$ = 0.9$\pm$0.5 and a surface flux of F$_X$ = (1.17$\pm$0.16) 10$^{-7}$ erg s$^{-1}$ cm$^{-2}$ sr$^{-1}$ in the 1--7 keV band. We estimated the contribution from unresolved point sources to the observed excess to be negligible. The observations suggest that a purely thermal origin of the emission is less likely than a non-thermal scenario. However, from simple modeling we cannot identify a clearly preferred scenario.Comment: 6 pages, 4 figures, accepted for publication by A&

Coronal emission from the shocked circumstellar ring of SN 1987A

High resolution spectra with UVES/VLT of SN 1987A from December 2000 until November 2005 show a number of high ionization lines from gas with velocities of roughly 350 km/s, emerging from the shocked gas formed by the ejecta-ring collision. These include coronal lines from [Fe X], [Fe XI] and [Fe XIV] which have increased by a factor of about 20 during the observed period. The evolution of the lines is similar to that of the soft X-rays, indicating that they arise in the same component. The line ratios are consistent with those expected from radiative shocks with velocity 310-390 km/s, corresponding to a shock temperature of (1.6-2.5) x 10^6 K. A fraction of the coronal emission may, however, originate in higher velocity adiabatic shocks.Comment: 11 pages, 10 figures, accepted for publication in A&

Mechanism of Cancer Cell Death Induced by Depletion of an Essential Replication Regulator

Background: Depletion of replication factors often causes cell death in cancer cells. Depletion of Cdc7, a kinase essential for initiation of DNA replication, induces cancer cell death regardless of its p53 status, but the precise pathways of cell death induction have not been characterized. Methodology/Principal Findings: We have used the recently-developed cell cycle indicator, Fucci, to precisely characterize the cell death process induced by Cdc7 depletion. We have also generated and utilized similar fluorescent cell cycle indicators using fusion with other cell cycle regulators to analyze modes of cell death in live cells in both p53-positive and-negative backgrounds. We show that distinct cell-cycle responses are induced in p53-positive and-negative cells by Cdc7 depletion. p53-negative cells predominantly arrest temporally in G2-phase, accumulating CyclinB1 and other mitotic regulators. Prolonged arrest at G2-phase and abrupt entry into aberrant M-phase in the presence of accumulated CyclinB1 are followed by cell death at the post-mitotic state. Abrogation of cytoplasmic CyclinB1 accumulation partially decreases cell death. The ATR-MK2 pathway is responsible for sequestration of CyclinB1 with 14-3-3s protein. In contrast, p53-positive cancer cells do not accumulate CyclinB1, but appear to die mostly through entry into aberrant S-phase after Cdc7 depletion. The combination of Cdc7 inhibition with known anti-cancer agents significantly stimulates cell death effects in cancer cells in a genotype-dependent manner, providing a strategic basis for future combination therapies

Collisional Ionization Equilibrium for Optically Thin Plasmas. I. Updated Recombination Rate Coefficients for Bare though Sodium-like Ions

Reliably interpreting spectra from electron-ionized cosmic plasmas requires accurate ionization balance calculations for the plasma in question. However, much of the atomic data needed for these calculations have not been generated using modern theoretical methods and are often highly suspect. This translates directly into the reliability of the collisional ionization equilibrium (CIE) calculations. We make use of state-of-the-art calculations of dielectronic recombination (DR) rate coefficients for the hydrogenic through Na-like ions of all elements from He up to and including Zn. We also make use of state-of-the-art radiative recombination (RR) rate coefficient calculations for the bare through Na-like ions of all elements from H through to Zn. Here we present improved CIE calculations for temperatures from $10^4$ to $10^9$ K using our data and the recommended electron impact ionization data of \citet{Mazz98a} for elements up to and including Ni and Mazzotta (private communication) for Cu and Zn. DR and RR data for ionization stages that have not been updated are also taken from these two additional sources. We compare our calculated fractional ionic abundances using these data with those presented by Mazzotta et al. for all elements from H to Ni. The differences in peak fractional abundance are up to 60%. We also compare with the fractional ionic abundances for Mg, Si, S, Ar, Ca, Fe, and Ni derived from the modern DR calculations of \citet{Gu03a,Gu04a} for the H-like through Na-like ions, and the RR calculations of \citet{Gu03b} for the bare through F-like ions. These results are in better agreement with our work, with differences in peak fractional abundance of less than 10%.Comment: 83 pages, 38 figures, 41 tables Accepted to ApJ

Replication Fork Reactivation in a dnaC2 Mutant at Non-Permissive Temperature in Escherichia coli

Replicative helicases unwind double-stranded DNA in front of the polymerase and ensure the processivity of DNA synthesis. In Escherichia coli, the helicase loader DnaC as well as factors involved in the formation of the open complex during the initiation of replication and primosomal proteins during the reactivation of arrested replication forks are required to recruit and deposit the replicative helicase onto single-stranded DNA prior to the formation of the replisome. dnaC2 is a thermosensitive allele of the gene specifying the helicase loader; at non-permissive temperature replication cannot initiate, but most ongoing rounds of replication continues through to completion (18% of dnaC2 cells fail to complete replication at non-permissive temperature). An assumption, which may be drawn from this observation, is that only a few replication forks are arrested under normal growth conditions. This assumption, however, is at odds with the severe and deleterious phenotypes associated with a null mutant of priA, the gene encoding a helicase implicated in the reactivation of arrested replication forks. We developed an assay that involves an abrupt inactivation of rounds of synchronized replication in a large population of cells, in order to evaluate the ability of dnaC2 cells to reactivate arrested replication forks at non-permissive temperature. We compared the rate at which arrested replication forks accumulated in dnaC2 priA+ and dnaC2 priA2 cells and observed that this rate was lower in dnaC2 priA+ cells. We conclude that while replication cannot initiate in a dnaC2 mutant at non-permissive temperature, a class of arrested replication forks (PriA-dependent and DnaC-independent) are reactivated within these cells

Characterization of Leishmania donovani MCM4: Expression Patterns and Interaction with PCNA

Events leading to origin firing and fork elongation in eukaryotes involve several proteins which are mostly conserved across the various eukaryotic species. Nuclear DNA replication in trypanosomatids has thus far remained a largely uninvestigated area. While several eukaryotic replication protein orthologs have been annotated, many are missing, suggesting that novel replication mechanisms may apply in this group of organisms. Here, we characterize the expression of Leishmania donovani MCM4, and find that while it broadly resembles other eukaryotes, noteworthy differences exist. MCM4 is constitutively nuclear, signifying that, unlike what is seen in S.cerevisiae, varying subcellular localization of MCM4 is not a mode of replication regulation in Leishmania. Overexpression of MCM4 in Leishmania promastigotes causes progress through S phase faster than usual, implicating a role for MCM4 in the modulation of cell cycle progression. We find for the first time in eukaryotes, an interaction between any of the proteins of the MCM2-7 (MCM4) and PCNA. MCM4 colocalizes with PCNA in S phase cells, in keeping with the MCM2-7 complex being involved not only in replication initiation, but fork elongation as well. Analysis of a LdMCM4 mutant indicates that MCM4 interacts with PCNA via the PIP box motif of MCM4 - perhaps as an integral component of the MCM2-7 complex, although we have no direct evidence that MCM4 harboring a PIP box mutation can still functionally associate with the other members of the MCM2-7 complex- and the PIP box motif is important for cell survival and viability. In Leishmania, MCM4 may possibly help in recruiting PCNA to chromatin, a role assigned to MCM10 in other eukaryotes

X-ray source populations in the Galactic Plane

We present the first results from the \xmm Galactic Plane Survey (XGPS). In the first phase of the programme, 22 pointings were used to cover a region of approximately three square degrees between 19\deg -- 22\deg in Galactic longitude and $\pm$0.6\deg in latitude. In total we have resolved over 400 point X-ray sources, at $\geq 5 \sigma$ significance, down to a flux limit of $\sim2 \times 10^{-14}$ \ergseccm (2--10 keV). The combination of the XGPS measurements in the hard X-ray band with the results from earlier surveys carried out by \asca and \chan reveals the form of the low-latitude X-ray source counts over 4 decades of flux. It appears that extragalactic sources dominate below $\sim10^{-13}$ \ergseccm (2--10 keV), with a predominantly Galactic source population present above this flux threshold. The nature of the faint Galactic population observed by \xmm remains uncertain, although cataclysmic variables and RS CVn systems may contribute substantially. \xmm observes an enhanced surface brightness in the Galactic plane in the 2--6 keV band associated with Galactic Ridge X-ray Emission (GRXE). The integrated contribution of Galactic sources plus the breakthrough of extragalactic signal accounts for up to 20 per cent of the observed surface brightness. The XGPS results are consistent with the picture suggested from a deep \chan observation in the Galactic plane, namely that the bulk of the GRXE is truly diffuse.Comment: 21 pages, 15 figures, accepted by MNRA

Interactions of the Human MCM-BP Protein with MCM Complex Components and Dbf4

MCM-BP was discovered as a protein that co-purified from human cells with MCM proteins 3 through 7; results which were recapitulated in frogs, yeast and plants. Evidence in all of these organisms supports an important role for MCM-BP in DNA replication, including contributions to MCM complex unloading. However the mechanisms by which MCM-BP functions and associates with MCM complexes are not well understood. Here we show that human MCM-BP is capable of interacting with individual MCM proteins 2 through 7 when co-expressed in insect cells and can greatly increase the recovery of some recombinant MCM proteins. Glycerol gradient sedimentation analysis indicated that MCM-BP interacts most strongly with MCM4 and MCM7. Similar gradient analyses of human cell lysates showed that only a small amount of MCM-BP overlapped with the migration of MCM complexes and that MCM complexes were disrupted by exogenous MCM-BP. In addition, large complexes containing MCM-BP and MCM proteins were detected at mid to late S phase, suggesting that the formation of specific MCM-BP complexes is cell cycle regulated. We also identified an interaction between MCM-BP and the Dbf4 regulatory component of the DDK kinase in both yeast 2-hybrid and insect cell co-expression assays, and this interaction was verified by co-immunoprecipitation of endogenous proteins from human cells. In vitro kinase assays showed that MCM-BP was not a substrate for DDK but could inhibit DDK phosphorylation of MCM4,6,7 within MCM4,6,7 or MCM2-7 complexes, with little effect on DDK phosphorylation of MCM2. Since DDK is known to activate DNA replication through phosphorylation of these MCM proteins, our results suggest that MCM-BP may affect DNA replication in part by regulating MCM phosphorylation by DDK

DnaC Inactivation in Escherichia coli K-12 Induces the SOS Response and Expression of Nucleotide Biosynthesis Genes

Background: Initiation of chromosome replication in E. coli requires the DnaA and DnaC proteins and conditionally-lethal dnaA and dnaC mutants are often used to synchronize cell populations. Methodology/Principal Findings: DNA microarrays were used to measure mRNA steady-state levels in initiation-deficient dnaA46 and dnaC2 bacteria at permissive and non-permissive temperatures and their expression profiles were compared to MG1655 wildtype cells. For both mutants there was altered expression of genes involved in nucleotide biosynthesis at the non-permissive temperature. Transcription of the dnaA and dnaC genes was increased at the non-permissive temperature in the respective mutant strains indicating auto-regulation of both genes. Induction of the SOS regulon was observed in dnaC2 cells at 38uC and 42uC. Flow cytometric analysis revealed that dnaC2 mutant cells at non-permissive temperature had completed the early stages of chromosome replication initiation. Conclusion/Significance: We suggest that in dnaC2 cells the SOS response is triggered by persistent open-complex formation at oriC and/or by arrested forks that require DnaC for replication restart