Targeted protein degradation is a powerful tool in determining the function of specific proteins or protein complexes. We fused nanobodies to SPOP, an adaptor protein of the Cullin-RING E3 ubiquitin ligase complex, resulting in rapid ubiquitination and subsequent proteasome-dependent degradation of specific nuclear proteins in mammalian cells and zebrafish embryos. This approach is easily modifiable, as substrate specificity is conferred by an antibody domain that can be adapted to target virtually any protein.

While transfer-RNAs (tRNAs) are known to transport amino acids to ribosome, new functions are being unveiled from tRNAs and their fragments beyond protein synthesis. Here we show that phosphorylation of 90-kDa RPS6K (ribosomal proteins S6 kinase) was enhanced by tRNALeu overexpression under amino acids starvation condition. The phosphorylation of 90-kDa RPS6K was decreased by siRNA specific to tRNALeu and was independent to mTOR (mammalian target of rapamycin) signaling. Among the 90-kDa RPS6K family, RSK1 (ribosomal S6 kinase 1) and MSK2 (mitogen-and stress-activated protein kinase 2) were the major kinases phosphorylated by tRNALeu overexpression. Through SILAC (stable isotope labeling by/with amino acids in cell culture) and combined mass spectrometry analysis, we identified EBP1 (ErbB3-binding protein 1) as the tRNALeu-binding protein. We suspected that the overexpression of free tRNALeu would reinforce ErbB2/ErbB3 signaling pathway by disturbing the interaction between ErbB3 and EBP1, resulting in RSK1/MSK2 phosphorylation, improving cell proliferation and resistance to death. Analysis of samples from patients with breast cancer also indicated an association between tRNALeu overexpression and the ErbB2-positive population. Our results suggested a possible link between tRNALeu overexpression and RSK1/MSK2 activation and ErbB2/ErbB3 signaling.

No abstract

The study is the result of an practical operation analysis for the full scale fishery product wastewater treatment plant with immersed MBR (iMBR) process. Since fishery product industries show a wide range of wastewater generation by the season, design and operation of the equalization basin are very important factor. The aeration system for the equalization basin mixing can save the chemical consumption for followed system through the restriction of acid fermentation. The concentrations of wastewater primary DAF process treated were BOD 2,291 mg/L, CODMn 530 mg/L, SS 256.8 mg/L, T-N 38 mg/L, T-P 13.5 mg/L respectively. It was considered that iMBR is the most efficient biological process for high salinity content wastewater since It is irrelevant to the capability of the sludge precipitation. SADp and SADm were 0.31, 26.5 m 3 /hr･m 3 respectively. In iMBR process, the critical F/M ratio was derived at 0.08~0.10 gBOD/gMLSS by analysing the correlations between MLSS, normalized TMP and temperature.