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Uracil occurs at replication forks via misincorporation of deoxyuridine monophosphate (dUMP) or via deamination of existing cytosines, which occurs 2-3 orders of magnitude faster in ssDNA than in dsDNA and is 100% miscoding. Tethering of UNG2 to proliferating cell nuclear antigen (PCNA) allows rapid post-replicative removal of misincorporated uracil, but potential 'pre-replicative' removal of deaminated cytosines in ssDNA has been questioned since this could mediate mutagenic translesion synthesis and induction of double-strand breaks. Here, we demonstrate that uracil-DNA glycosylase (UNG), but not SMUG1 efficiently excises uracil from replication protein A (RPA)-coated ssDNA and that this depends on functional interaction between the flexible winged-helix (WH) domain of RPA2 and the N-terminal RPA-binding helix in UNG. This functional interaction is promoted by mono-ubiquitination and diminished by cell-cycle regulated phosphorylations on UNG. Six other human proteins bind the RPA2-WH domain, all of which are involved in DNA repair and replication fork remodelling. Based on this and the recent discovery of the AP site crosslinking protein HMCES, we propose an integrated model in which templated repair of uracil and potentially other mutagenic base lesions in ssDNA at the replication fork, is orchestrated by RPA. The UNG:RPA2-WH interaction may also play a role in adaptive immunity by promoting efficient excision of AID-induced uracils in transcribed immunoglobulin loci.

While Brexit has encouraged further interest in the phenomenon of right-wing Euroscepticism, it also serves as an opportunity to investigate pro-European attitudes. The Brexit process put pro-Europeans in a position where their place within the integration project was under immediate threat and required something so unusual as pro-European mobilisation. This thesis aims to contribute to that investigation by examining the discussion of the EEA- and Norway-plus mod-els featured online by the British newspaper the Guardian. Specifically, it considers the debate in terms of evaluations that explicitly promotes or rejects these models as desirable future rela-tionships. The analysis treats these evaluations as proxies for European integration to identify how the British centre-left valuate and evaluate the integration project. The decision to focus on the cen-tre-left follows from the British left’s historical relationship to European integration as well as its position somewhat on the side of the Brexit debate. This context is thought to make the public discourse on the centre-left a most-likely case for new or alternative modes of justification and defence of the integration project. The analysis finds considerable congruence between the discussion in the Guardian and British interpretations of European integration more generally, as well as with the Conservative-dominated referendum debate. Specifically, supporters typically based their position on an un-derstanding of integration as an economic project, while detractors usually emphasise sovereign-ty and democracy. The congruence suggests that, despite different conclusion about the desira-bility of European integration, a consistent understanding existed in Britain across time and ide-ology platform, of what integration is ‘supposed’ to be. There is little evidence of innovation in the Brexit discourse on the centre-left; pre-existing frames and interpretations remained un-touched.

Additional file 6: Table S2. Proteotypic PRM peptides.

As of June 2020, the number of people infected with severe acute respiratory coronavirus 2 (SARS-CoV-2) continues to skyrocket, with more than 6.7 million cases worldwide. Both the World Health Organization (WHO) and United Nations (UN) has highlighted the need for better control of SARS-CoV-2 infections. However, developing novel virus-specific vaccines, monoclonal antibodies and antiviral drugs against SARS-CoV-2 can be time-consuming and costly. Convalescent sera and safe-in-man broad-spectrum antivirals (BSAAs) are readily available treatment options. Here, we developed a neutralization assay using SARS-CoV-2 strain and Vero-E6 cells. We identified the most potent sera from recovered patients for the treatment of SARS-CoV-2-infected patients. We also screened 136 safe-in-man broad-spectrum antivirals against the SARS-CoV-2 infection in Vero-E6 cells and identified nelfinavir, salinomycin, amodiaquine, obatoclax, emetine and homoharringtonine. We found that a combination of orally available virus-directed nelfinavir and host-directed amodiaquine exhibited the highest synergy. Finally, we developed a website to disseminate the knowledge on available and emerging treatments of COVID-19. This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.