title: Modulating the Structure and Function of an Aminoacyl-tRNA Synthetase Cofactor by Biotinylation.
abstract: Arc1p is a yeast-specific tRNA-binding protein that forms a ternary complex with glutamyl-tRNA synthetase (GluRSc) and methionyl-tRNA synthetase (MetRS) in the cytoplasm to regulate their catalytic activities and subcellular distributions. Despite Arc1p not being involved in any known biotin-dependent reaction, it is a natural target of biotin modification. Results presented herein show that biotin modification had no obvious effect on the growth-supporting activity, subcellular distribution, tRNA binding, or interactions of Arc1p with GluRSc and MetRS. Nevertheless, biotinylation of Arc1p was temperature dependent; raising the growth temperature from 30 to 37 °C drastically reduced its biotinylation level. As a result, Arc1p purified from a yeast culture that had been grown overnight at 37 °C was essentially biotin free. Non-biotinylated Arc1p was more heat stable, more flexible in structure, and more effective than its biotinylated counterpart in promoting glutamylation activity of the otherwise inactive GluRSc at 37 °C in vitro Our study suggests that the structure and function of Arc1p can be modulated via biotinylation in response to temperature changes.
authors.affiliation: From the Department of Life Sciences and.
authors.forename: Chih-Yao
authors.identifier: 0000-0002-6688-1520
authors.initials: CY
authors.lastname: Chang
authors.affiliation: From the Department of Life Sciences and.
authors.forename: Chia-Pei
authors.identifier: 0000-0002-1412-9652
authors.initials: CP
authors.lastname: Chang
authors.affiliation: the Department of Biotechnology, University of Calcutta, Kolkata 700019, India, and.
authors.forename: Shruti
authors.initials: S
authors.lastname: Chakraborty
authors.affiliation: the Division of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan Town, Miaoli 35053, Taiwan.
authors.forename: Shao-Win
authors.initials: SW
authors.lastname: Wang
authors.affiliation: the Graduate Institute of Statistics, National Central University, Jungli District, Taoyuan 32001, Taiwan.
authors.forename: Yi-Kuan
authors.initials: YK
authors.lastname: Tseng
authors.affiliation: From the Department of Life Sciences and dukewang@cc.ncu.edu.tw.
authors.forename: Chien-Chia
authors.identifier: 0000-0002-7104-1307
authors.initials: CC
authors.lastname: Wang
mesh_terms: D019921:Biotinylation; D005975:Glutamate-tRNA Ligase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D008718:Methionine-tRNA Ligase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055550:Protein Stability; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C102145:ARC1 protein, S cerevisiae; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D008718:Methionine-tRNA Ligase; D005975:Glutamate-tRNA Ligase
keywords: aminoacyl tRNA synthetase; biotin; post-translational modification (PTM); protein synthesis; transfer RNA (tRNA)