Human cytomegalovirus immediate-early 1 protein rewires upstream STAT3 to downstream STAT1 signaling switching an IL6-type to an IFNγ-like response

MN and CP were supported by the Wellcome Trust (www.wellcome.ac.uk) Institutional Strategic Support Fund and CP was supported by the Deutsche Forschungsgemeinschaft (PA 815/2-1; www.dfg.de).The human cytomegalovirus (hCMV) major immediate-early 1 protein (IE1) is best known for activating transcription to facilitate viral replication. Here we present transcriptome data indicating that IE1 is as significant a repressor as it is an activator of host gene expression. Human cells induced to express IE1 exhibit global repression of IL6- and oncostatin M-responsive STAT3 target genes. This repression is followed by STAT1 phosphorylation and activation of STAT1 target genes normally induced by IFNγ. The observed repression and subsequent activation are both mediated through the same region (amino acids 410 to 445) in the C-terminal domain of IE1, and this region serves as a binding site for STAT3. Depletion of STAT3 phenocopies the STAT1-dependent IFNγ-like response to IE1. In contrast, depletion of the IL6 receptor (IL6ST) or the STAT kinase JAK1 prevents this response. Accordingly, treatment with IL6 leads to prolonged STAT1 instead of STAT3 activation in wild-type IE1 expressing cells, but not in cells expressing a mutant protein (IE1dl410-420) deficient for STAT3 binding. A very similar STAT1-directed response to IL6 is also present in cells infected with a wild-type or revertant hCMV, but not an IE1dl410-420 mutant virus, and this response results in restricted viral replication. We conclude that IE1 is sufficient and necessary to rewire upstream IL6-type to downstream IFNγ-like signaling, two pathways linked to opposing actions, resulting in repressed STAT3- and activated STAT1-responsive genes. These findings relate transcriptional repressor and activator functions of IE1 and suggest unexpected outcomes relevant to viral pathogenesis in response to cytokines or growth factors that signal through the IL6ST-JAK1-STAT3 axis in hCMV-infected cells. Our results also reveal that IE1, a protein considered to be a key activator of the hCMV productive cycle, has an unanticipated role in tempering viral replication.Publisher PDFPeer reviewe

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

An analytic proof system for common knowledge logic over S5

In this paper we present an analytic proof system for multi-modal logic with common-knowledge over S5 (called S5-CKL). The system is an annotated cyclic calculus manipulating two-sided Gentzen sequents and extending a known system for multi-modalS5. First a direct argument is used to show that the system is sound. Using a canonical model construction, we then show that the system is analytically complete. In particular, the use of the cut-rule is restricted to analytic cuts. Exploiting this analyticity, we then reduce the provability problem of a given sequent to the problem of solving a certain parity game. As a consequence we obtain an optimal decision procedure for proof search and thereby for the validity problem of S5-CKL

A Family of Decidable Bi-intuitionistic Modal Logics

We investigate intuitionistic logics extended with both the co-implication connective of Hilbert–Brouwer logic and with diamond and box modalities. We use a Kripke semantics based on frames with two ‘forth’ confluence conditions on the modal relation with respect to the intuitionistic relation. We give sound and strongly complete axiomatisations for entailment on this class of frames, and give similar axiomatisations for the subclasses of frames satisfying any combination of reflexivity, transitivity, and seriality. We then prove that all of these logics are decidable, by proving that they have the finite frame property

Ill-Founded Proof Systems for Intuitionistic Linear-Time Temporal Logic

We introduce ill-founded sequent calculi for two intuitionistic linear-time temporal logics. Both logics are based on the language of intuitionistic propositional logic with ‘next’and ‘until’operators and are evaluated on dynamic Kripke models wherein the intuitionistic and temporal accessibility relations are assumed to satisfy one of two natural confluence properties: forward confluence in one case, and both forward and backward confluence in the other. The presented sequent calculi are cut-free and incorporate a simple form of formula nesting. Soundness of the calculi is shown by a standard argument and completeness via proof search

Response of peripheral arterial pulse wave velocity to acute exercise in patients after recent myocardial infarction and healthy controls

Background Many studies found increased central arterial stiffness and poor endothelial function in patients with coronary artery disease (CAD). Acute exercise has been shown to decrease peripheral pulse wave velocity (pPWV) in young healthy volunteers. We hypothesized the response to acute exercise to be diminished in CAD patients compared to healthy young (HY) and age-matched (HAM) controls. Methods In 21 patients after recent myocardial infarction (CAD), 11 HAM and 10 HY pPWV was measured by applanation tonometry at the proximal femoral artery and the posterior tibial artery at rest and from 5 to 15 min after cessation of exhaustive exercise. Heart rate (HR) and blood pressure (BP) were monitored continuously. Resting central PWV (cPWV) was measured between the carotid and femoral arteries. Resting values and reponses to acute exercise were compared between the three groups and predictors for pPWV response were sought. Results The response in pPWV to acute exercise seen in HY (lowering in pPWV by 17%) was absent in both CAD and HAM. Resting pPWV was not statistically different between the three groups, while cPWV was comparable in CAD and HAM but 17% lower in HY. Predictors for response in pPWV to exercise were age (Spearman r = 0.48), cPWV (r = 0.34) and response in diastolic BP (r = 0.32). Conclusion The response in pPWV to acute exercise observed in HY was absent in CAD and HAM. In dilated peripheral arteries, PWV may reflect stiffness of passive vessel structures, which are likely to increase with age in healthy persons and CAD alike.ISSN:1932-620

Ingenuity analysis¹ of human genes repressed by IE1 and activated by STAT3, IL6 or OSM.

<p>Ingenuity analysis<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005748#t001fn001" target="_blank">¹</a> of human genes repressed by IE1 and activated by STAT3, IL6 or OSM.</p

Residues within IE1 region 410–445 are required for phosphorylation of STAT1 and up-regulation of IFNγ-responsive genes.

<p>(A) TetR cells without (w/o) or with inducible expression of the indicated HA-tagged wild-type or mutant IE1 proteins were treated with dox for 72 h. Whole cell protein extracts were prepared and analyzed by immunoblotting for IE1 (HA tag), pSTAT1 (Y701), pSTAT1 (S727), total STAT1 and GAPDH. (B) TetR cells without (w/o) or with inducible expression of the indicated HA-tagged wild-type or mutant IE1 proteins were treated with dox for 72 h. Relative mRNA expression levels were determined by RT-qPCR with primers specific for the STAT1 target genes CXCL10 and CXCL11. Results were normalized to TUBB, and means and standard deviations of two biological and two technical replicates are shown in comparison to IE1-negative TetR cells (set to 1).</p

Systematic deletion analysis of C-terminal IE1 residues 373–491.

<p>(A) Schematic overview of amino acids 373–491 in the tested wild-type and mutant IE1 proteins. Positions of the low-complexity motifs (acidic domains AD1-3 and serine/proline-rich region S/P), the SUMOylation site (K450) and the chromatin tethering domain (CTD) are shown. (B) TetR cells without (w/o) or with inducible expression of the indicated HA-tagged wild-type or mutant IE1 proteins were treated with dox for 72 h. Whole cell protein extracts were prepared and analyzed by immunoblotting for IE1 (HA tag) and GAPDH. (C) TetR cells without (w/o) or with inducible expression of the indicated HA-tagged wild-type or mutant IE1 proteins were treated with dox for 72 h. Whole cell extracts prepared in the presence of N-ethylmaleimide were used for immunoprecipitation with anti-HA-agarose, and samples were analyzed by immunoblotting for IE1 (HA tag) and SUMO1.</p

IE1 switches an IL6-type to an IFNγ-like response.

<p>(A) TetR (w/o) or TetR-IE1 (IE1) cells were treated with dox for 72 h and with IL6 plus IL6R (IL6/Rα) for the indicated times. Whole cell protein extracts were prepared and analyzed by immunoblotting for IE1, total STAT1, pSTAT1 (Y701), total STAT3α, pSTAT3 (Y705) and GAPDH. (B) TetR (w/o) or TetR-IE1 cells expressing HA-tagged wild-type IE1 or IE1dl410-420 were treated with dox for 72 h and with solvent, IFNα, IFNγ or IL6 plus IL6R (IL6/Rα) for 24 h. Whole cell protein extracts were analyzed by immunoblotting for IE1, total STAT1, pSTAT1 (Y701) and GAPDH. (C) TetR (w/o) or TetR-IE1 cells expressing HA-tagged wild-type IE1 or IE1dl410-420 were treated with dox for 72 h and with solvent, IFNα, IFNγ or IL6 plus IL6R (IL6/Rα) for 24 h. Relative mRNA levels were determined by RT-qPCR for the type I IFN/STAT2 target genes OAS1 and EIF2AK2 (protein kinase R) (left panels), the type II IFN/STAT1 target genes CXCL10 and CXCL11 (middle panels) and the IL6/STAT3 target genes CXCL12 and SOCS3 (right panels). Results were normalized to TUBB, and means and standard deviations of biological triplicates are shown in comparison to solvent-treated TetR cells (set to 1).</p