The spliceosome is a dynamic ribonucleoprotein (RNP) machine that catalyzes the removal of introns in the two transesterification steps of eukaryotic pre-mRNA splicing. Here we used single molecule fluorescence resonance energy transfer to monitor the distance of the 5′ splice site (5′SS) and branchpoint (BP) of pre-mRNA in affinity-purified spliceosomes stalled by a mutation in the DExD/H-box helicase Prp2 immediately prior to the first splicing step. Addition of recombinant Prp2 together with NTP and protein cofactor Spp2 rearranges the spliceosome-substrate complex to reversibly explore conformations with proximal 5′SS and BP that accommodate chemistry. Addition of Cwc25 then strongly biases this equilibrium towards the proximal conformation, promoting efficient first-step splicing. The spliceosome thus functions as a biased Brownian ratchet machine where a helicase unlocks thermal fluctuations subsequently rectified by a cofactor “pawl”, a principle possibly widespread among the many helicase-driven RNPs.

BackgroundAlthough it is well established that heavy alcohol consumption increases the risk of hypertension, the risk associated with low levels of alcohol intake in men and women is unclear.Methods and ResultsWe searched Medline and Embase for original cohort studies on the association between average alcohol consumption and incidence of hypertension in people without hypertension. Random‐effects meta‐analyses and metaregressions were conducted. Data from 20 articles with 361 254 participants (125 907 men and 235 347 women) and 90 160 incident cases of hypertension (32 426 men and 57 734 women) were included. In people drinking 1 to 2 drinks/day (12 g of pure ethanol per drink), incidence of hypertension differed between men and women (relative riskwomen vs men=0.79; 95% confidence interval, 0.67–0.93). In men, the risk for hypertension in comparison with abstainers was relative risk=1.19 (1.07–1.31; I2=59%), 1.51 (1.30–1.76), and 1.74 (1.35–2.24) for consumption of 1 to 2, 3 to 4, and 5 or more standard drinks per day, respectively. In women, there was no increased risk for 1 to 2 drinks/day (relative risk=0.94; 0.88–1.01; I2=73%), and an increased risk for consumption beyond this level (relative risk=1.42; 1.22–1.66).ConclusionsAny alcohol consumption was associated with an increase in the risk for hypertension in men. In women, there was no risk increase for consumption of 1 to 2 drinks/day and an increased risk for higher consumption levels. We did not find evidence for a protective effect of alcohol consumption in women, contrary to earlier meta‐analyses.

Oncolytic viruses (OV) are an emerging class of anticancer bio-therapeutics that induce antitumor immunity through selective replication in tumor cells. However, the efficacy of OVs as single agents remains limited. We introduce a strategy that boosts the therapeutic efficacy of OVs by combining their activity with immuno-modulating, small molecule protein tyrosine phosphatase inhibitors. We report that vanadium-based phosphatase inhibitors enhance OV infection in vitro and ex vivo, in resistant tumor cell lines. Furthermore, vanadium compounds increase antitumor efficacy in combination with OV in several syngeneic tumor models, leading to systemic and durable responses, even in models otherwise refractory to OV and drug alone. Mechanistically, this involves subverting the antiviral type I IFN response toward a death-inducing and pro-inflammatory type II IFN response, leading to improved OV spread, increased bystander killing of cancer cells, and enhanced antitumor immune stimulation. Overall, we showcase a new ability of vanadium compounds to simultaneously maximize viral oncolysis and systemic anticancer immunity, offering new avenues for the development of improved immunotherapy strategies.