Severe course with lethal hepatocellular injury and skeletal muscular dysgenesis in a neonate with infantile liver failure syndrome type 1 caused by novel LARS1 mutations.

Infantile liver failure syndrome type 1 (ILFS1) is a recently recognized autosomal recessive disorder caused by deleterious mutations in the leucyl-tRNA synthetase 1 gene (LARS1). The LARS1 enzyme is responsible for incorporation of the amino acid leucine during protein polypeptide synthesis. Individuals with LARS1 mutations typically show liver failure from infancy to early childhood during periods of illness or other physiological stress. While 25 patients from 15 families with ILFS1 have been reported in the literature, histological reports from autopsy findings are limited. We report here a premature male neonate who presented with severe intrauterine growth retardation, microcytic anemia, and fulminant liver failure, and who was a compound heterozygote for two novel deleterious mutations in LARS1. An autopsy showed fulminant hepatitis-like hepatocellular injury and fibrogenesis in the liver and a lack of uniformity in skeletal muscle, accompanied by the disruption of striated muscle fibers. Striking dysgenesis in skeletal muscle detected in the present case indicates the effect of LARS1 functional deficiency on the musculature. Whole-exome sequencing may be useful for neonates with unexplained early liver failure if extensive genetic and metabolic testing is inconclusive.

Leucyl-tRNA synthetase deficiency systemically induces excessive autophagy in zebrafish.

Leucyl-tRNA synthetase (LARS) is an enzyme that catalyses the ligation of leucine with leucine tRNA. LARS is also essential to sensitize the intracellular leucine concentration to the mammalian target of rapamycin complex 1 (mTORC1) activation. Biallelic mutation in the LARS gene causes infantile liver failure syndrome type 1 (ILFS1), which is characterized by acute liver failure, anaemia, and neurological disorders, including microcephaly and seizures. However, the molecular mechanism underlying ILFS1 under LARS deficiency has been elusive. Here, we generated Lars deficient (larsb-/-) zebrafish that showed progressive liver failure and anaemia, resulting in early lethality within 12 days post fertilization. The atg5-morpholino knockdown and bafilomycin treatment partially improved the size of the liver and survival rate in larsb-/- zebrafish. These findings indicate the involvement of autophagy in the pathogenesis of larsb-/- zebrafish. Indeed, excessive autophagy activation was observed in larsb-/- zebrafish. Therefore, our data clarify a mechanistic link between LARS and autophagy in vivo. Furthermore, autophagy regulation by LARS could lead to development of new therapeutics for IFLS1.

Unique residues crucial for optimal editing in yeast cytoplasmic Leucyl-tRNA synthetase are revealed by using a novel knockout yeast strain.

Leucyl-tRNA synthetase (LeuRS) contains an editing domain that discriminates leucine from noncognate amino acids to ensure translational fidelity. In this study, a knock-out strain for Saccharomyces cerevisiae LeuRS was constructed to analyze in vivo the tRNA aminoacylation properties of S. cerevisiae and human cytoplasmic LeuRSs. The activities of several editing-defective mutants of ycLeuRS were determined in vitro and compared with those obtained in vivo in a complementation assay performed in the knock-out strain. The editing activities of these mutants were analyzed in the presence of either norvaline, a leucine analogue, or AN2690, a specific inhibitor that targets the editing active site. In general, the in vivo data are consistent with those obtained in vitro. Our results show that ycLeuRS post-transfer editing plays a crucial role in the establishment of the aminoacylation fidelity. When impaired, the viability of cells bearing editing-defective mutants is drastically decreased in the presence of noncognate amino acid. This study also emphasizes the crucial function of some semi-conserved residues around the editing site in modulating the editing efficiency. The assay system can be used to test the effect of compounds that potentially target the aminoacylation or editing active site of fungal LeuRS.