Shuffled ATG8 interacting motifs form an ancestral bridge between UFMylation and autophagy

UFMylation involves the covalent modification of substrate proteins with UFM1 (Ubiquitin‐fold modifier 1) and is important for maintaining ER homeostasis. Stalled translation triggers the UFMylation of ER‐bound ribosomes and activates C53‐mediated autophagy to clear toxic polypeptides. C53 contains noncanonical shuffled ATG8‐interacting motifs (sAIMs) that are essential for ATG8 interaction and autophagy initiation. However, the mechanistic basis of sAIM‐mediated ATG8 interaction remains unknown. Here, we show that C53 and sAIMs are conserved across eukaryotes but secondarily lost in fungi and various algal lineages. Biochemical assays showed that the unicellular alga Chlamydomonas reinhardtii has a functional UFMylation pathway, refuting the assumption that UFMylation is linked to multicellularity. Comparative structural analyses revealed that both UFM1 and ATG8 bind sAIMs in C53, but in a distinct way. Conversion of sAIMs into canonical AIMs impaired binding of C53 to UFM1, while strengthening ATG8 binding. Increased ATG8 binding led to the autoactivation of the C53 pathway and sensitization of Arabidopsis thaliana to ER stress. Altogether, our findings reveal an ancestral role of sAIMs in UFMylation‐dependent fine‐tuning of C53‐mediated autophagy activation

An Experimental Investigation of Ethanol-Diesel Blends on Performance and Exhaust Emissions of Diesel Engines

Ethanol is a promising alternative fuel, due to its renewable biobased origin. Also, it has lower carbon content than diesel fuel and it is oxygenated. For this reason, ethanol is providing remarkable potential to reduce particulate emulsions in compression-ignition engines. In this study, performance of ethanol-diesel blends has been investigated experimentally. Tested fuels were mineral diesel fuel (E0D100), 15% (v/v) ethanol/diesel fuel blend (E15D85), and 30% (v/v) ethanol/diesel fuel blend (E30D70). Firstly, the solubility of ethanol and diesel was experienced. Engine tests were carried out to reveal the performance and emissions of the engine fuelled with the blends. Full load operating conditions at various engine speeds were investigated. Engine brake torque, brake power, brake specific fuel consumption, brake thermal efficiency, exhaust gas temperature, and finally exhaust emissions were measured. Performance of the tested engine decreased substantially while improvement on smoke and gaseous emissions makes ethanol blend favorable

Effect of hydrogen–diesel dual-fuel usage on performance, emissions and diesel combustion in diesel engines

Diesel engines are inevitable parts of our daily life and will be in the future. Expensive after-treatment technologies to fulfil normative legislations about the harmful tail-pipe emissions and fuel price increase in recent years created expectations from researchers for alternative fuel applications on diesel engines. This study investigates hydrogen as additive fuel in diesel engines. Hydrogen was introduced into intake manifold using gas injectors as additive fuel in gaseous form and also diesel fuel was injected into cylinder by diesel injector and used as igniter. Energy content of introduced hydrogen was set to 0%, 25% and 50% of total fuel energy, where the 0% references neat diesel operation without hydrogen injection. Test conditions were set to full load at 750, 900, 1100, 1400, 1750 and finally 2100 r/min engine speed. Variation in engine performance, emissions and combustion characteristics with hydrogen addition was investigated. Hydrogen introduction into the engine by 25% and 50% of total charge energy reveals significant decrease in smoke emissions while dramatic increase in nitrogen oxides. With increasing hydrogen content, a slight rise is observed in total unburned hydrocarbons although CO 2 and CO gaseous emissions reduced considerably. Maximum in-cylinder gas pressure and rate of heat release peak values raised with hydrogen fraction

Effect of the use of natural gas–diesel fuel mixture on performance, emissions, and combustion characteristics of a compression ignition engine

A compression ignition engine with a mechanical fuel system was converted into common rail fuel system by means of a self-developed electronic control unit. The engine was modified to be operated with mixtures of diesel and natural gas fuels in dual-fuel mode. Then, diesel fuel was injected into the cylinder while natural gas was injected into intake manifold with both injectors controlled with the electronic control unit. Energy content of the sprayed gas fuel was varied in the amounts of 0% (only diesel fuel), 15%, 40%, and 75% of total fuel’s energy content. All tests were carried out at constant engine speed of 1500 r/min at full load. In addition to the experiments, the engine was modeled with a one-dimensional commercial software. The experimental and numerical results were compared and found to be in reasonable agreement with each other. Both NO x and soot emissions were dropped with 15% and 40%, respectively, energy content rates in gas–fuel mixture compared to only diesel fuel. However, an increase was observed in carbon monoxide emissions with 15% natural gas fuel addition compared to only diesel fuel. Although smoke emission was reduced with natural gas fuel addition, there was a dramatic increase in NO x emissions with 75% natural gas fuel addition

Shuffled ATG8 interacting motifs form an ancestral bridge between UFMylation and C53-mediated autophagy

UFMylation mediates the covalent modification of substrate proteins with UFM1 (Ubiquitin-fold modifier 1) and regulates the selective degradation of endoplasmic reticulum (ER) via autophagy (ER-phagy) to maintain ER homeostasis. Specifically, collisions of the ER-bound ribosomes trigger ribosome UFMylation, which in turn activates C53-mediated autophagy that clears the toxic incomplete polypeptides. C53 has evolved non-canonical shuffled ATG8 interacting motifs (sAIMs) that are essential for ATG8 interaction and autophagy initiation. Why these non-canonical motifs were selected during evolution, instead of canonical ATG8 interacting motifs remains unknown. Here, using a phylogenomics approach, we show that UFMylation is conserved across the eukaryotes and secondarily lost in fungi and some other species. Further biochemical assays have confirmed those results and showed that the unicellular algae, Chlamydomonas reinhardtii has a functional UFMylation machinery, overturning the assumption that this process is linked to multicellularity. Our conservation analysis also revealed that UFM1 co-evolves with the sAIMs in C53, reflecting a functional link between UFM1 and the sAIMs. Using biochemical and structural approaches, we confirmed the interaction of UFM1 with the C53 sAIMs and found that UFM1 and ATG8 bound to the sAIMs in a different mode. Conversion of sAIMs into canonical AIMs prevented binding of UFM1 to C53, while strengthening ATG8 interaction. This led to the autoactivation of the C53 pathway and sensitized Arabidopsis thaliana to ER stress. Altogether, our findings reveal an ancestral toggle switch embodied in the sAIMs that regulates C53-mediated autophagy to maintain ER homeostasis.Competing Interest StatementThe authors have declared no competing interest