Transfer-RNA-mediated enhancement of ribosomal proteins S6 kinases signalling for cell proliferation

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 tRNA^Leu overexpression under amino acids starvation condition. The phosphorylation of 90-kDa RPS6K was decreased by siRNA specific to tRNA^Leu and was independent to mTOR (mammalian target of rapamycin) signalling. 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 tRNA^Leu overexpression. Through SILAC (stable isotope labelling by/with amino acids in cell culture) and combined mass spectrometry analysis, we identified EBP1 (ErbB3-binding protein 1) as the tRNA^Leu-binding protein. We suspected that the overexpression of free tRNA^Leu would reinforce ErbB2/ErbB3 signalling 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 tRNA^Leu overexpression and the ErbB2-positive population. Our results suggested a possible link between tRNA^Leu overexpression and RSK1/MSK2 activation and ErbB2/ErbB3 signalling.