MARCH6 and TRC8 facilitate the quality control of cytosolic and tail‐anchored proteins

Misfolded or damaged proteins are typically targeted for destruction by proteasome‐mediated degradation, but the mammalian ubiquitin machinery involved is incompletely understood. Here, using forward genetic screens in human cells, we find that the proteasome‐mediated degradation of the soluble misfolded reporter, mCherry‐CL1, involves two ER‐resident E3 ligases, MARCH6 and TRC8. mCherry‐CL1 degradation is routed via the ER membrane and dependent on the hydrophobicity of the substrate, with complete stabilisation only observed in double knockout MARCH6/TRC8 cells. To identify a more physiological correlate, we used quantitative mass spectrometry and found that TRC8 and MARCH6 depletion altered the turnover of the tail‐anchored protein heme oxygenase‐1 (HO‐1). These E3 ligases associate with the intramembrane cleaving signal peptide peptidase (SPP) and facilitate the degradation of HO‐1 following intramembrane proteolysis. Our results highlight how ER‐resident ligases may target the same substrates, but work independently of each other, to optimise the protein quality control of selected soluble and tail‐anchored proteins.     

Are the benefits of yield responses to nitrogen fertilizer application in the bioenergy crop Miscanthus × giganteus offset by increased soil emissions of nitrous oxide?

A field trial was carried out on a 15 year old Miscanthus stand, subject to nitrogen fertilizer treatments of 0, 63 and 125 kg‐N ha^?1, measuring N₂O emissions, as well as annual crop yield over a full year. N₂O emission intensity (N₂O emissions calculated as a function of above‐ground biomass) was significantly affected by fertilizer application, with values of 52.2 and 59.4 g N₂O‐N t^?1 observed at 63 and 125 kg‐N ha^?1, respectively, compared to 31.3 g N₂O‐N t^?1 in the zero fertilizer control. A life cycle analyses approach was applied to calculate the increase in yield required to offset N₂O emissions from Miscanthus through fossil fuel substitution in the fuel chain. For the conditions observed during the field trial yield increases of 0.33 and 0.39 t ha^?1 were found to be required to offset N₂O emissions from the 63 kg‐N ha^?1 treatment, when replacing peat and coal, respectively, while increases of 0.71 and 0.83 t ha^?1 were required for the 125 kg‐N ha^?1 treatment, for each fuel. These values are considerably less than the mean above‐ground biomass yield increases observed here of 1.57 and 2.79 t ha^?1 at fertilization rates 63 and 125 kg‐N ha^?1 respectively. Extending this analysis to include a range of fertilizer application rates and N₂O emission factors found increases in yield necessary to offset soil N₂O emissions ranging from 0.26 to 2.54 t ha^?1. These relatively low yield increase requirements indicate that where nitrogen fertilizer application improves yield, the benefits of such a response will not be offset by soil N₂O emissions.