TY - JOUR
T1 - Substrate interaction defects in histidyl-tRNA synthetase linked to dominant axonal peripheral neuropathy
AU - Abbott, Jamie A.
AU - Meyer-Schuman, Rebecca
AU - Lupo, Vincenzo
AU - Feely, Shawna
AU - Mademan, Inès
AU - Oprescu, Stephanie N.
AU - Griffin, Laurie B.
AU - Alberti, M. Antonia
AU - Casasnovas, Carlos
AU - Aharoni, Sharon
AU - Basel-Vanagaite, Lina
AU - Züchner, Stephan
AU - De Jonghe, Peter
AU - Baets, Jonathan
AU - Shy, Michael E.
AU - Espinós, Carmen
AU - Demeler, Borries
AU - Antonellis, Anthony
AU - Francklyn, Christopher
N1 - Funding Information:
and Blood Institute, Grant/Award Number: T32 HL 007594-30; Research Fund-Flanders (FWO); National Institute of General Medical Sciences, Grant/Award Numbers: GM 007315, GM 007544, GM 07863, GM118647, GM54899; Michigan Pre-doctoral Training in Genetics Program, Grant/Award Number: GM007544; Innovation by Science and Technology (IWT); National Institutes of Health Cellular and Molecular Biology Training Grant, Grant/Award Number: GM007315; National Institiues of Health F30 NRSA, Grant/Award Number: NS092238; Miguel Servet Program, Grant/Award Number: CPII14/00002; Muscular Dystrophy Association, Grant/Award Number: MDA294479; Aide à la Recherche ASBL; EU FP7/2007-2013, Grant/Award Number: 2012–305121; San Antonio Cancer Institute Grant
PY - 2018/3
Y1 - 2018/3
N2 - Histidyl-tRNA synthetase (HARS) ligates histidine to cognate tRNA molecules, which is required for protein translation. Mutations in HARS cause the dominant axonal peripheral neuropathy Charcot-Marie-Tooth disease type 2W (CMT2W); however, the precise molecular mechanism remains undefined. Here, we investigated three HARS missense mutations associated with CMT2W (p.Tyr330Cys, p.Ser356Asn, and p.Val155Gly). The three mutations localize to the HARS catalytic domain and failed to complement deletion of the yeast ortholog (HTS1). Enzyme kinetics, differential scanning fluorimetry (DSF), and analytical ultracentrifugation (AUC) were employed to assess the effect of these substitutions on primary aminoacylation function and overall dimeric structure. Notably, the p.Tyr330Cys, p.Ser356Asn, and p.Val155Gly HARS substitutions all led to reduced aminoacylation, providing a direct connection between CMT2W-linked HARS mutations and loss of canonical ARS function. While DSF assays revealed that only one of the variants (p.Val155Gly) was less thermally stable relative to wild-type, all three HARS mutants formed stable dimers, as measured by AUC. Our work represents the first biochemical analysis of CMT-associated HARS mutations and underscores how loss of the primary aminoacylation function can contribute to disease pathology.
AB - Histidyl-tRNA synthetase (HARS) ligates histidine to cognate tRNA molecules, which is required for protein translation. Mutations in HARS cause the dominant axonal peripheral neuropathy Charcot-Marie-Tooth disease type 2W (CMT2W); however, the precise molecular mechanism remains undefined. Here, we investigated three HARS missense mutations associated with CMT2W (p.Tyr330Cys, p.Ser356Asn, and p.Val155Gly). The three mutations localize to the HARS catalytic domain and failed to complement deletion of the yeast ortholog (HTS1). Enzyme kinetics, differential scanning fluorimetry (DSF), and analytical ultracentrifugation (AUC) were employed to assess the effect of these substitutions on primary aminoacylation function and overall dimeric structure. Notably, the p.Tyr330Cys, p.Ser356Asn, and p.Val155Gly HARS substitutions all led to reduced aminoacylation, providing a direct connection between CMT2W-linked HARS mutations and loss of canonical ARS function. While DSF assays revealed that only one of the variants (p.Val155Gly) was less thermally stable relative to wild-type, all three HARS mutants formed stable dimers, as measured by AUC. Our work represents the first biochemical analysis of CMT-associated HARS mutations and underscores how loss of the primary aminoacylation function can contribute to disease pathology.
KW - Charcot-Marie-Tooth disease type 2W
KW - aminoacyl-tRNA synthetase
KW - hereditary motor and sensory neuropathy
KW - histidyl-tRNA synthetase
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U2 - 10.1002/humu.23380
DO - 10.1002/humu.23380
M3 - Article
C2 - 29235198
AN - SCOPUS:85038939838
VL - 39
SP - 415
EP - 432
JO - Human Mutation
JF - Human Mutation
SN - 1059-7794
IS - 3
ER -