Substrate interaction defects in histidyl-tRNA synthetase linked to dominant axonal peripheral neuropathy
Por:
J. ABBOTT, R. MEYER-SCHUMAN, V. LUPO, S. FEELY, I. MADEMAN, S. OPRESCU, L. GRIFFIN, M. ALBERTI, C. CASASNOVAS, S. AHARONI, L. BASEL-VANAGAITE, S. ZUCHNER, P. DE JONGHE, J. BAETS, M. SHY, C. ESPINOS, B. DEMELER, A. ANTONELLIS and C. FRANCKLYN
Publicada:
1 mar 2018
Resumen:
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.
Filiaciones:
J. ABBOTT:
Univ Vermont, Coll Med, Dept Biochem, Burlington, VT 05405 USA
R. MEYER-SCHUMAN:
Univ Michigan, Dept Human Genet, Med Sch, Ann Arbor, MI 48109 USA
:
CIPF, Unit Genet & Genom Neuromuscular Disorders, Valencia, Spain
S. FEELY:
Univ Iowa Hosp & Clin, Dept Neurol, Iowa City, IA 52242 USA
I. MADEMAN:
VIB, Ctr Mol Neurol, Neurogenet Grp, Antwerp, Belgium
Univ Antwerp, Inst Born Bunge, Lab Neuromuscular Pathol, Antwerp, Belgium
S. OPRESCU:
Univ Michigan, Dept Human Genet, Med Sch, Ann Arbor, MI 48109 USA
L. GRIFFIN:
Univ Michigan, Med Sch, Cellular & Mol Biol Program, Ann Arbor, MI 48109 USA
Univ Michigan, Med Sch, Med Scientist Training Program, Ann Arbor, MI 48109 USA
M. ALBERTI:
Hosp Univ Bellvitge, Dept Neurol, Barcelona, Spain
C. CASASNOVAS:
Hosp Univ Bellvitge, Dept Neurol, Barcelona, Spain
S. AHARONI:
Tel Aviv Univ, Sackler Fac Med, Schneider Childrens Med Ctr Israel, Inst Child Neurol, Tel Aviv, Israel
L. BASEL-VANAGAITE:
Tel Aviv Univ, Sackler Fac Med, Tel Aviv, Israel
Rabin Med Ctr, Raphael Recanati Genet Inst, Beilinson Campus, Petah Tiqwa, Israel
Schneider Childrens Med Ctr, Pediat Genet Unit, Petah Tiqwa, Israel
Rabin Med Ctr, Felsenstein Med Res Ctr, Petah Tiqwa, Israel
S. ZUCHNER:
Univ Miami, Miller Sch Med, Dept Human Genet, Dr John TMcDonald Fdn, Miami, FL 33136 USA
Univ Miami, Miller Sch Med, John P Hussman Inst Human Genom, Miami, FL 33136 USA
P. DE JONGHE:
VIB, Ctr Mol Neurol, Neurogenet Grp, Antwerp, Belgium
Univ Antwerp, Inst Born Bunge, Lab Neuromuscular Pathol, Antwerp, Belgium
Antwerp Univ Hosp, Dept Neurol, Antwerp, Belgium
J. BAETS:
VIB, Ctr Mol Neurol, Neurogenet Grp, Antwerp, Belgium
Univ Antwerp, Inst Born Bunge, Lab Neuromuscular Pathol, Antwerp, Belgium
Antwerp Univ Hosp, Dept Neurol, Antwerp, Belgium
M. SHY:
Univ Iowa Hosp & Clin, Dept Neurol, Iowa City, IA 52242 USA
:
CIPF, Unit Genet & Genom Neuromuscular Disorders, Valencia, Spain
B. DEMELER:
Univ Texas Hlth Sci Ctr San Antonio, Dept Biochem, San Antonio, TX 78229 USA
A. ANTONELLIS:
Univ Michigan, Dept Human Genet, Med Sch, Ann Arbor, MI 48109 USA
C. FRANCKLYN:
Univ Vermont, Coll Med, Dept Biochem, Burlington, VT 05405 USA
Green Submitted, Green Accepted, gold
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