@article{dcc25c44d8ee480796e1f2e09c5b896c,
title = "Leucine-sensing mechanism of leucyl-tRNA synthetase 1 for mTORC1 activation",
abstract = "Leucyl-tRNA synthetase 1 (LARS1) mediates activation of leucine-dependent mechanistic target of rapamycin complex 1 (mTORC1) as well as ligation of leucine to its cognate tRNAs, yet its mechanism of leucine sensing is poorly understood. Here we describe leucine binding-induced conformational changes of LARS1. We determine different crystal structures of LARS1 complexed with leucine, ATP, and a reaction intermediate analog, leucyl-sulfamoyl-adenylate (Leu-AMS), and find two distinct functional states of LARS1 for mTORC1 activation. Upon leucine binding to the synthetic site, H251 and R517 in the connective polypeptide and 50FPYPY54 in the catalytic domain change the hydrogen bond network, leading to conformational change in the C-terminal domain, correlating with RagD association. Leucine binding to LARS1 is increased in the presence of ATP, further augmenting leucine-dependent interaction of LARS1 and RagD. Thus, this work unveils the structural basis for leucine-dependent long-range communication between the catalytic and RagD-binding domains of LARS1 for mTORC1 activation.",
keywords = "X-ray crystallography, conformational change, leucine sensing, leucyl-tRNA synthetase 1, mechanistic target of rapamycin complex 1",
author = "Sulhee Kim and Ina Yoon and Jonghyeon Son and Junga Park and Kibum Kim and Lee, {Ji Ho} and Park, {Sam Yong} and Kang, {Beom Sik} and Han, {Jung Min} and Hwang, {Kwang Yeon} and Sunghoon Kim",
note = "Funding Information: This work was supported by the Global Frontier Project grant (NRF-M3A6A4-2010-0029785 and 2013M3A6A4044795) of the National Research Foundation (NRF) funded by the Ministry of Science and ICT (MSIT) of Korea, the Yonsei University Research Fund (2020-22-0358), and an NRF grant funded by the Korean government (2018M3A9F3055925, 2020R1A2C2005670, 2020R1I1A1A01067423, 2020R1A2C2099586, and 2020M3E5E2040282). We thank the staff at the PLS 5C, 7A, 11C beamline in South Korea, Photon Factory 1A, and Spring-8 44XU beamline in Japan for the use of their excellent facilities and assistance with X-ray data collection. We also acknowledge the Korean Basic Science Institute (Daejeon, Korea) for the use of a circular dichroism spectrophotometer. Sulhee Kim was supported by the WISET program (2018-644) and NRF-2019R1I1A1A01056 of Korea. K.Y.H. and Sunghoon Kim designed all experiments. Sulhee Kim cloned, purified, and crystallized WT of LARS1s. J.S. and Sulhee Kim prepared LARS1 mutants. J.S. and Sulhee Kim collected and processed the diffraction data of LARS1s. Sulhee Kim, I.Y. J.P. K.K. J.M.H. J.S. and J.-H.L. performed the biochemical analysis. S.-Y.P. B.S.K. and K.Y.H. determined the crystal structures of LARS1s. I.Y. J.S. Sulhee Kim, Sunghoon Kim, and K.Y.H. analyzed the data and wrote the manuscript. The authors declare no competing interests. Funding Information: This work was supported by the Global Frontier Project grant (NRF-M3A6A4-2010-0029785 and 2013M3A6A4044795) of the National Research Foundation (NRF) funded by the Ministry of Science and ICT (MSIT) of Korea, the Yonsei University Research Fund ( 2020-22-0358 ), and an NRF grant funded by the Korean government ( 2018M3A9F3055925 , 2020R1A2C2005670 , 2020R1I1A1A01067423 , 2020R1A2C2099586 , and 2020M3E5E2040282 ). We thank the staff at the PLS 5C, 7A, 11C beamline in South Korea, Photon Factory 1A, and Spring-8 44XU beamline in Japan for the use of their excellent facilities and assistance with X-ray data collection. We also acknowledge the Korean Basic Science Institute (Daejeon, Korea) for the use of a circular dichroism spectrophotometer. Sulhee Kim was supported by the WISET program ( 2018-644 ) and NRF-2019R1I1A1A01056 of Korea. Publisher Copyright: {\textcopyright} 2021 The Authors",
year = "2021",
month = apr,
day = "27",
doi = "10.1016/j.celrep.2021.109031",
language = "English",
volume = "35",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "4",
}