Early Nuclear Events after Herpesviral Infection

Herpesviruses are important pathogens that can cause significant morbidity and mortality in the human population. Herpesviruses have a double-stranded DNA genome, and viral genome replication takes place inside the nucleus. Upon entering the nucleus, herpesviruses have to overcome the obstacle of cellular proteins in order to enable viral gene expression and genome replication. In this review, we want to highlight cellular proteins that sense incoming viral genomes of the DNA-damage repair (DDR) pathway and of PML-nuclear bodies (PML-NBs) that all can act as antiviral restriction factors within the first hours after the viral genome is released into the nucleus. We show the function and significance of both nuclear DNA sensors, the DDR and PML-NBs, and demonstrate for three human herpesviruses of the alpha-, beta- and gamma-subfamilies, HSV-1, HCMV and KSHV respectively, how viral tegument proteins antagonize these pathways

Episomal replication timing of γ-herpesviruses in latently infected cells

AbstractThis study addresses the timing of gammaherpesviral episomal DNA replication with respect to the cell cycle. For the first time we analyzed a rhadinovirus, the prototype Herpesvirus saimiri (HVS), and compared it to the lymphocryptovirus Epstein–Barr virus (EBV). Newly synthesized DNA of latently infected B- or T-cells was first BrdU-labeled; then we sorted the cells corresponding to cell cycle phases G0/1, G2/M, and S (4 fractions S1–S4) and performed anti-BrdU chromatin immunoprecipitation. Next, DNA of different viral gene loci was quantitatively detected together with cellular control genes of known replication time. The sensitive technique is further enhanced by an internal coprecipitation standard for increased precision. Both gammaherpesviruses replicated very early in S-phase, together with cellular euchromatin. Our work suggests that early S-phase DNA replication is a general characteristic of episomal herpesviral genomes

The Clustering of Galaxies in the SDSS-III Baryon Oscillation Spectroscopic Survey: Signs of Neutrino Mass in Current Cosmological Data Sets

We investigate the cosmological implications of the latest growth of structure measurement from the Baryon Oscillation Spectroscopic Survey (BOSS) CMASS Data Release 11 with particular focus on the sum of the neutrino masses, Σmv. We examine the robustness of the cosmological constraints from the baryon acoustic oscillation (BAO) scale, the Alcock-Paczynski effect and redshift-space distortions (DV/rs, FAP, fσ8) of Beutler et al., when introducing a neutrino mass in the power spectrum template. We then discuss how the neutrino mass relaxes discrepancies between the cosmic microwave background (CMB) and other low-redshift measurements within Λ cold dark matter. Combining our cosmological constraints with 9-year Wilkinson Microwave Anisotropy Probe (WMAP9) yields Σmv = 0.36 ± 0.14 eV (68 per cent c.l.), which represents a 2.6σ preference for non-zero neutrino mass. The significance can be increased to 3.3σ when including weak lensing results and other BAO constraints, yielding Σmv = 0.35 ± 0.10 eV (68 per cent c.l.). However, combining CMASS with Planck data reduces the preference for neutrino mass to ~2σ. When removing the CMB lensing effect in the Planck temperature power spectrum (by marginalizing over AL, we see shifts of ~1σ in σ8 and Ωm, which have a significant effect on the neutrino mass constraints. In the case of CMASS plus Planck without the AL lensing signal, we find a preference for a neutrino mass of Σmv = 0.34 ± 0.14 eV (68 per cent c.l.), in excellent agreement with the WMAP9+CMASS value. The constraint can be tightened to 3.4σ yielding Σmv = 0.36 ± 0.10 eV (68 per cent c.l.) when weak lensing data and other BAO constraints are included

The Clustering of Galaxies in the SDSS-III Baryon Oscillation Spectroscopic Survey: Testing Gravity with Redshift Space Distortions using the Power Spectrum Multipoles

We analyse the anisotropic clustering of the Baryon Oscillation Spectroscopic Survey (BOSS) CMASS Data Release 11 (DR11) sample, which consists of 690 827 galaxies in the redshift range 0.43 \u3c z \u3c 0.7 and has a sky coverage of 8498 deg2. We perform our analysis in Fourier space using a power spectrum estimator suggested by Yamamoto et al. We measure the multipole power spectra in a self-consistent manner for the first time in the sense that we provide a proper way to treat the survey window function and the integral constraint, without the commonly used assumption of an isotropic power spectrum and without the need to split the survey into subregions. The main cosmological signals exploited in our analysis are the baryon acoustic oscillations and the signal of redshift space distortions, both of which are distorted by the Alcock-Paczynski effect. Together, these signals allow us to constrain the distance ratio DV(zeff)/rs(zd) = 13.89 ± 0.18, the Alcock-Paczynski parameter FAP(zeff) = 0.679 ± 0.031 and the growth rate of structure f(zeff)σ8(zeff) = 0.419 ± 0.044 at the effective redshift zeff = 0.57. We emphasize that our constraints are robust against possible systematic uncertainties. In order to ensure this, we perform a detailed systematics study against CMASS mock galaxy catalogues and N-body simulations. We find that such systematics will lead to 3.1 per cent uncertainty for σ8 if we limit our fitting range to k = 0.01-0.20 h Mpc-1, where the statistical uncertainty is expected to be three times larger. We did not find significant systematic uncertainties for DV/rs or FAP. Combining our data set with Planck to test General Relativity (GR) through the simple γ-parametrization, where the growth rate is given by f (z) = Ωγm m(z), reveals a ~2σ tension between the data and the prediction by GR. The tension between our result and GR can be traced back to a tension in the clustering amplitude σ8 between CMASS and Planck

The Clustering of Galaxies in the Completed SDSS-III Baryon Oscillation Spectroscopic Survey: Observational Systematics and Baryon Acoustic Oscillations in the Correlation Function

We present baryon acoustic oscillation (BAO) scale measurements determined from the clustering of 1.2 million massive galaxies with redshifts 0.2 \u3c z \u3c 0.75 distributed over 9300 deg2, as quantified by their redshift-space correlation function. In order to facilitate these measurements, we define, describe, and motivate the selection function for galaxies in the final data release (DR12) of the SDSS III Baryon Oscillation Spectroscopic Survey (BOSS). This includes the observational footprint, masks for image quality and Galactic extinction, and weights to account for density relationships intrinsic to the imaging and spectroscopic portions of the survey. We simulate the observed systematic trends in mock galaxy samples and demonstrate that they impart no bias on BAO scale measurements and have a minor impact on the recovered statistical uncertainty. We measure transverse and radial BAO distance measurements in 0.2 \u3c z \u3c 0.5, 0.5 \u3c z \u3c 0.75, and (overlapping) 0.4 \u3c z \u3c 0.6 redshift bins. In each redshift bin, we obtain a precision that is 2.7 per cent or better on the radial distance and 1.6 per cent or better on the transverse distance. The combination of the redshift bins represents 1.8 per cent precision on the radial distance and 1.1 per cent precision on the transverse distance. This paper is part of a set that analyses the final galaxy clustering data set from BOSS. The measurements and likelihoods presented here are combined with others in Alam et al. to produce the final cosmological constraints from BOSS

Redirecting T Cells to Ewing's Sarcoma Family of Tumors by a Chimeric NKG2D Receptor Expressed by Lentiviral Transduction or mRNA Transfection

We explored the possibility to target Ewing's sarcoma family of tumors (ESFT) by redirecting T cells. To this aim, we considered NKG2D-ligands (NKG2D-Ls) as possible target antigens. Detailed analysis of the expression of MICA, MICB, ULBP-1, -2, and -3 in fourteen ESFT cell lines revealed consistent expression of at least one NKG2D-L. Thus, for redirecting T cells, we fused a CD3ζ/CD28-derived signaling domain to the ectodomain of NKG2D, however, opposite transmembrane orientation of this signaling domain and NKG2D required inverse orientation fusion of either of them. We hypothesized that the particularly located C-terminus of the NKG2D ectodomain should allow reengineering of the membrane anchoring from a native N-terminal to an artificial C-terminal linkage. Indeed, the resulting chimeric NKG2D receptor (chNKG2D) was functional and efficiently mediated ESFT cell death triggered by activated T cells. Notably, ESFT cells with even low NKG2D-L expression were killed by CD8pos and also CD4pos cells. Both, mRNA transfection and lentiviral transduction resulted in high level surface expression of chNKG2D. However, upon target-cell recognition receptor surface levels were maintained by tranfected RNA only during the first couple of hours after transfection. Later, target-cell contact resulted in strong and irreversible receptor down-modulation, whereas lentivirally mediated expression of chNKG2D remained constant under these conditions. Together, our study defines NKG2D-Ls as targets for a CAR-mediated T cell based immunotherapy of ESFT. A comparison of two different methods of gene transfer reveals strong differences in the susceptibility to ligand-induced receptor down-modulation with possible implications for the applicability of RNA transfection

Gammaherpesviral Tegument Proteins, PML-Nuclear Bodies and the Ubiquitin-Proteasome System

Gammaherpesviruses like Epstein-Barr virus (EBV) and Kaposi’s sarcoma-associated herpesvirus (KSHV) subvert the ubiquitin proteasome system for their own benefit in order to facilitate viral gene expression and replication. In particular, viral tegument proteins that share sequence homology to the formylglycineamide ribonucleotide amidotransferase (FGARAT, or PFAS), an enzyme in the cellular purine biosynthesis, are important for disrupting the intrinsic antiviral response associated with Promyelocytic Leukemia (PML) protein-associated nuclear bodies (PML-NBs) by proteasome-dependent and independent mechanisms. In addition, all herpesviruses encode for a potent ubiquitin protease that can efficiently remove ubiquitin chains from proteins and thereby interfere with several different cellular pathways. In this review, we discuss mechanisms and functional consequences of virus-induced ubiquitination and deubiquitination for early events in gammaherpesviral infection

Gammaherpesviral Tegument Proteins, PML-Nuclear Bodies and the Ubiquitin-Proteasome System

Gammaherpesviruses like Epstein-Barr virus (EBV) and Kaposi’s sarcoma-associated herpesvirus (KSHV) subvert the ubiquitin proteasome system for their own benefit in order to facilitate viral gene expression and replication. In particular, viral tegument proteins that share sequence homology to the formylglycineamide ribonucleotide amidotransferase (FGARAT, or PFAS), an enzyme in the cellular purine biosynthesis, are important for disrupting the intrinsic antiviral response associated with Promyelocytic Leukemia (PML) protein-associated nuclear bodies (PML-NBs) by proteasome-dependent and independent mechanisms. In addition, all herpesviruses encode for a potent ubiquitin protease that can efficiently remove ubiquitin chains from proteins and thereby interfere with several different cellular pathways. In this review, we discuss mechanisms and functional consequences of virus-induced ubiquitination and deubiquitination for early events in gammaherpesviral infection