Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human Lysyl-tRNA Synthetase

被引:8
作者
Bin Kwon, Soon [1 ]
Yu, Ji Eun [1 ]
Park, Chan [1 ]
Lee, Jiseop [1 ]
Seong, Baik L. [1 ]
机构
[1] Yonsei Univ, Dept Biotechnol, Coll Life Sci & Biotechnol, Seoul 03722, South Korea
基金
新加坡国家研究基金会;
关键词
aminoacyl-tRNA synthetase; LysRS; hRID; disorder-helix transition; ESCHERICHIA-COLI; UNSTRUCTURED PROTEINS; NONCANONICAL ROLES; ENTROPIC BRISTLES; BINDING DOMAIN; ENERGY CONTENT; TRANSLATION; AGGREGATION; DOWNSTREAM; SOLUBILITY;
D O I
10.3390/ijms19103016
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Eukaryotic lysyl-tRNA synthetases (LysRS) have an N-terminal appended tRNA-interaction domain (RID) that is absent in their prokaryotic counterparts. This domain is intrinsically disordered and lacks stable structures. The disorder-to-order transition is induced by tRNA binding and has implications on folding and subsequent assembly into multi-tRNA synthetase complexes. Here, we expressed and purified RID from human LysRS (hRID) in Escherichia coli and performed a detailed mutagenesis of the appended domain. hRID was co-purified with nucleic acids during Ni-affinity purification, and cumulative mutations on critical amino acid residues abolished RNA binding. Furthermore, we identified a structural ensemble between disordered and helical structures in non-RNA-binding mutants and an equilibrium shift for wild-type into the helical conformation upon RNA binding. Since mutations that disrupted RNA binding led to an increase in non-functional soluble aggregates, a stabilized RNA-mediated structural transition of the N-terminal appended domain may have implications on the functional organization of human LysRS and multi-tRNA synthetase complexes in vivo.
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页数:15
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