Substrate interaction defects in histidyl-tRNA synthetase linked to dominant axonal peripheral neuropathy

Jamie A. Abbott, Rebecca Meyer-Schuman, Vincenzo Lupo, Shawna Feely, Inès Mademan, Stephanie N. Oprescu, Laurie B. Griffin, M. Antonia Alberti, Carlos Casasnovas, Sharon Aharoni, Lina Basel-Vanagaite, Stephan L Zuchner, Peter De Jonghe, Jonathan Baets, Michael E. Shy, Carmen Espinós, Borries Demeler, Anthony Antonellis, Christopher Francklyn

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
JournalHuman Mutation
DOIs
StateAccepted/In press - Jan 1 2017

Fingerprint

Histidine-tRNA Ligase
Peripheral Nervous System Diseases
Aminoacylation
Charcot-Marie-Tooth Disease
Fluorometry
Mutation
Ultracentrifugation
Protein Biosynthesis
Missense Mutation
Transfer RNA
Histidine
Catalytic Domain
Yeasts
Pathology
Enzymes

Keywords

  • Aminoacyl-tRNA synthetase
  • Charcot-Marie-Tooth disease type 2W
  • Hereditary motor and sensory neuropathy
  • Histidyl-tRNA synthetase

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Abbott, J. A., Meyer-Schuman, R., Lupo, V., Feely, S., Mademan, I., Oprescu, S. N., ... Francklyn, C. (Accepted/In press). Substrate interaction defects in histidyl-tRNA synthetase linked to dominant axonal peripheral neuropathy. Human Mutation. https://doi.org/10.1002/humu.23380

Substrate interaction defects in histidyl-tRNA synthetase linked to dominant axonal peripheral neuropathy. / Abbott, Jamie A.; Meyer-Schuman, Rebecca; Lupo, Vincenzo; Feely, Shawna; Mademan, Inès; Oprescu, Stephanie N.; Griffin, Laurie B.; Alberti, M. Antonia; Casasnovas, Carlos; Aharoni, Sharon; Basel-Vanagaite, Lina; Zuchner, Stephan L; De Jonghe, Peter; Baets, Jonathan; Shy, Michael E.; Espinós, Carmen; Demeler, Borries; Antonellis, Anthony; Francklyn, Christopher.

In: Human Mutation, 01.01.2017.

Research output: Contribution to journalArticle

Abbott, JA, Meyer-Schuman, R, Lupo, V, Feely, S, Mademan, I, Oprescu, SN, Griffin, LB, Alberti, MA, Casasnovas, C, Aharoni, S, Basel-Vanagaite, L, Zuchner, SL, De Jonghe, P, Baets, J, Shy, ME, Espinós, C, Demeler, B, Antonellis, A & Francklyn, C 2017, 'Substrate interaction defects in histidyl-tRNA synthetase linked to dominant axonal peripheral neuropathy', Human Mutation. https://doi.org/10.1002/humu.23380
Abbott, Jamie A. ; Meyer-Schuman, Rebecca ; Lupo, Vincenzo ; Feely, Shawna ; Mademan, Inès ; Oprescu, Stephanie N. ; Griffin, Laurie B. ; Alberti, M. Antonia ; Casasnovas, Carlos ; Aharoni, Sharon ; Basel-Vanagaite, Lina ; Zuchner, Stephan L ; De Jonghe, Peter ; Baets, Jonathan ; Shy, Michael E. ; Espinós, Carmen ; Demeler, Borries ; Antonellis, Anthony ; Francklyn, Christopher. / Substrate interaction defects in histidyl-tRNA synthetase linked to dominant axonal peripheral neuropathy. In: Human Mutation. 2017.
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abstract = "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.",
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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 - Zuchner, Stephan L

AU - De Jonghe, Peter

AU - Baets, Jonathan

AU - Shy, Michael E.

AU - Espinós, Carmen

AU - Demeler, Borries

AU - Antonellis, Anthony

AU - Francklyn, Christopher

PY - 2017/1/1

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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.

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