Leucine-sensing mechanism of leucyl-tRNA synthetase 1 for mTORC1 activation

Kim, Sulhee; Yoon, Ina; Son, Jonghyeon; Park, Junga; Kim, Ki-Bum; Lee, Ji Ho; Park, Sam Yong; Kang, Beom Sik; Han, Jung Min; Hwang, Kwang Yeon; Kim, Sung Hoon

doi:10.1016/j.celrep.2021.109031

Cited by 27 publications

(24 citation statements)

References 57 publications

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“…LARS1 WT binds to leucine, whereas LARS1 Y52A/Y54A/H91A does not bind to leucine. Figure reprinted with permission from Kim et al. (2021) .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…The extracted T i values are listed in Figure 3 . Figure reprinted with permission from Kim et al. (2021) .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…In this step, we obtain crystals of LARS1-Leu syn , LARS1-ATP syn , LARS1-Leu-AMS syn , and LARS1 methyl -Leu-AMS syn ( Kim et al., 2021 ). And then, we performed the post-crystallization soaking and cooling in cryoprotectants at −20°C ( Rould et al., 1991 ; Heras and Martin, 2005 ).…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…

Figure 1 Overall Domain arrangement of LARS1 CD, catalytic domain; LSD, leucine-specific domain; CP, connective polypeptide; SC, stem-contact; CTD, C-terminal domain; RBD, RagD-binding domain; UNE-L, unnamed domain of LARS. Figure reprinted with permission from Kim et al. (2021) .…”

Section: Before You Beginmentioning

confidence: 99%

“…Collectively, this allowed us to build the RagD binding domain, which was shown to be a dynamic region of LARS1 refractory to crystallization. For complete details on the use and execution of this protocol, please refer to Kim et al. (2021) .…”

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confidence: 99%

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Protocol for improving diffraction quality of leucyl-tRNA synthetase 1 with methylation and post-crystallization soaking and cooling in cryoprotectants

et al. 2021

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Summary Leucyl-tRNA synthetase 1 (LARS1) synthesizes Leu-tRNA Leu for protein synthesis and plays an important role in mTORC1 activation by sensing intracellular leucine concentrations. Here, we describe a protocol for the purification, reductive methylation, binding affinity measurement by microscale thermophoresis, T i value measurement by Tycho, and post-crystallization soaking and cooling in cryoprotectants to improve crystallization of LARS1. Collectively, this allowed us to build the RagD binding domain, which was shown to be a dynamic region of LARS1 refractory to crystallization. For complete details on the use and execution of this protocol, please refer to Kim et al. (2021) .

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“…LARS1 WT binds to leucine, whereas LARS1 Y52A/Y54A/H91A does not bind to leucine. Figure reprinted with permission from Kim et al. (2021) .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…The extracted T i values are listed in Figure 3 . Figure reprinted with permission from Kim et al. (2021) .…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…

Section: Before You Beginmentioning

confidence: 99%

mentioning

confidence: 99%

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Protocol for improving diffraction quality of leucyl-tRNA synthetase 1 with methylation and post-crystallization soaking and cooling in cryoprotectants

et al. 2021

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Mitochondrial‐Derived Peptide MOTS‐c Suppresses Ovarian Cancer Progression by Attenuating USP7‐Mediated LARS1 Deubiquitination

Yin,

Li,

et al. 2024

Advanced Science

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Mitochondrial‐nuclear communication plays a vital role in maintaining cellular homeostasis. MOTS‐c, a short peptide derived from the 12S rRNA of mitochondrial DNA, has been suggested as a retrograde mitochondrial signal. Although recent clinical studies have suggested a possible link between MOTS‐c and human cancer, the role of MOTS‐c in tumorigenesis has yet to be investigated. Here, MOTS‐c levels are found to be reduced in both serum and tumor tissues from ovarian cancer (OC) patients, which are associated with poor patients’ prognosis. Exogenous MOTS‐c inhibits the proliferation, migration and invasion of OC cells, and induces cell cycle arrest and apoptosis. Mechanistically, MOTS‐c interacts with LARS1 and promotes its ubiquitination and proteasomal degradation. In addition, USP7 was identified as a deubiquitinase of LARS1, and MOTS‐c can attenuates USP7‐mediated LARS1 deubiquitination by competing with USP7 for binding to LARS1. Besides, LARS1 was found to be increased and play an important oncogenic function in OC. More importantly, MOTS‐c displays a marked anti‐tumor effect on OC growth without systemic toxicity in vivo. In conclusion, this study reveals a crucial role of MOTS‐c in OC and provides a possibility for MOTS‐c as a therapeutic target for the treatment of this manlignacy.

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Two siblings with acute necrotizing encephalopathy associated with variants of LARS1

Uehara,

Seki,

Nonoda

et al. 2024

American J of Med Genetics Pt A

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Acute necrotizing encephalopathy (ANE) is a rapidly progressive encephalopathy of unknown etiology. The underlying mechanisms are highly heterogeneous, often including genetic backgrounds. Variants of LARS1, encoding the leucyl‐tRNA synthetase 1, are responsible for infantile liver failure syndrome 1. We describe two siblings with ANE caused by compound heterozygous variants of LARS1. Patient 1 was a 17‐month‐old girl. She presented with generalized seizure and liver dysfunction due to influenza type A infection. Brain magnetic resonance imaging on day 4 of onset showed diffuse high‐intensity signals consistent with ANE. She died on day 10. Patient 2, a younger male sibling of patient 1, had mild to moderate developmental delay and growth failure at the age of 18 months. He showed a markedly elevated level of transaminases triggered by infection with human herpesvirus 6. On day 4 of onset, he had generalized seizures. Brain computed tomography showed a diffuse symmetrical hypodensity consistent with ANE. He died on day 7. Whole exome sequencing identified the compound heterozygous variants in LARS1 (NM_020117.11) as c.83_88delinsAATGGGATA, p.(Arg28_Phe30delinsLysTryAspIle) and c.1283C>T, p.(Pro428Leu) in both siblings. The severe neurologic phenotype, found in our patients, reflects the complicated pathogenesis of LARS1‐related disorder.

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Leucine-sensing mechanism of leucyl-tRNA synthetase 1 for mTORC1 activation

Cited by 27 publications

References 57 publications

Protocol for improving diffraction quality of leucyl-tRNA synthetase 1 with methylation and post-crystallization soaking and cooling in cryoprotectants

Protocol for improving diffraction quality of leucyl-tRNA synthetase 1 with methylation and post-crystallization soaking and cooling in cryoprotectants

Mitochondrial‐Derived Peptide MOTS‐c Suppresses Ovarian Cancer Progression by Attenuating USP7‐Mediated LARS1 Deubiquitination

Two siblings with acute necrotizing encephalopathy associated with variants of LARS1

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