Dataset Information

Phenotypic rescue of a novel heterozygous HARS L400F variant associated with Charcot-Marie-Tooth disease

ABSTRACT: Cytoplasmic histidyl-tRNA synthetase (HARS1) is an essential protein in translation ligating histidine to its cognate tRNAHis. HARS1 is one of several aminoacyl-tRNA synthetase associated with Charcot-Marie-Tooth Disease, an axonal peripheral neuropathy. Advances in genetic testing identify many variants of uncertain significance. We characterize a novel heterozygous allele in HARS1, c.1200G>T (pLeu400Phe) with a familial inheritance pattern of peripheral neuropathy. Using a humanized yeast model and biochemical assays, we determined that L400F causes protein aggregation in the recombinant protein and in the humanized yeast model. L400F leads to a pronounced growth defect, especially at elevated temperatures, that persists in a heterozygous humanized yeast model, indicating a dominant negative mechanism. Cognate substrate supplementation does not ameliorate the growth defect. L400F leads to the activation of the integrated stress response (ISR) and upregulated chaperone expression in yeast. The yeast growth phenotype can be rescued by inhibition of the ISR using a specific inhibitor of the ISR activating protein general control non-depressible 2 (GCN2).

INSTRUMENT(S):

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Kyle Hoffman

LAB HEAD: Ilka Heinemann

PROVIDER: PXD067079 | Pride | 2026-03-02

REPOSITORIES: Pride

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			Action	DRS
	S24017-L400f3-20240731.mgf	Mgf
	S24017-L400f3-20240731.raw	Raw
	S24017-L400f3-20240731.raw.mgf	Mgf
	S24017-L400f3-20240731.raw.mzxml	Mzxml
	S24017_L400f1_20240725.mgf	Mgf

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Publications

Inhibition of the integrated stress response rescues a histidyl-tRNA synthetase variant associated with Charcot-Marie-Tooth disease.

Mahmood Maria M Wilhelm Sarah D P SDP Mendes Marisa I MI Smith Desiree E DE Kenana Rosan R Hoffman Kyle K Moresco Angelica A AA Siu Victoria Mok VM Heinemann Ilka U IU

NAR molecular medicine 20260101 1

Cytoplasmic histidyl-tRNA synthetase (HARS1) is an essential protein in translation, ligating histidine to its cognate tRNA<sup>His</sup>. HARS1 is one of several aminoacyl-transfer RNA (tRNA) synthetases associated with Charcot-Marie-Tooth disease, an axonal peripheral neuropathy. Advances in genetic testing identify many variants of uncertain significance. We characterize a novel heterozygous allele in HARS1, c.1200G > T (p.Leu400Phe) with a familial inheritance pattern of peripheral neuropath ...[more]

PMID: 41737028

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