Simos G, Segref A, Fasiolo F, Hellmuth K, Shevchenko A, Mann M, Hurt E C
University of Heidelberg, Institut für Biochemie I, Germany.
EMBO J. 1996 Oct 1;15(19):5437-48.
Arc1p was found in a screen for components that interact genetically with Los1p, a nuclear pore-associated yeast protein involved in tRNA biogenesis. Arc1p is associated with two proteins which were identified as methionyl-tRNA and glutamyl-tRNA synthetase (MetRS and GluRS) by a new mass spectrometry method. ARC1 gene disruption leads to slow growth and reduced MetRS activity, and synthetically lethal arc1- mutants are complemented by the genes for MetRS and GluRS. Recombinant Arc1p binds in vitro to purified monomeric yeast MetRS, but not to an N-terminal truncated form, and strongly increases its apparent affinity for tRNAMet. Furthermore, Arc1p, which is allelic to the quadruplex nucleic acid binding protein G4p1, exhibits specific binding to tRNA as determined by gel retardation and UV-cross-linking. Arc1p is, therefore, a yeast protein with dual specificity: it associates with tRNA and aminoacyl-tRNA synthetases. This functional interaction may be required for efficient aminoacylation in vivo.
在一项针对与Los1p发生遗传相互作用的成分的筛选中发现了Arc1p，Los1p是一种与核孔相关的酵母蛋白，参与tRNA生物合成。通过一种新的质谱方法，Arc1p与两种蛋白质相关联，这两种蛋白质被鉴定为甲硫氨酰-tRNA合成酶和谷氨酰胺-tRNA合成酶（MetRS和GluRS）。ARC1基因破坏导致生长缓慢和MetRS活性降低，并且合成致死的arc1-突变体可被MetRS和GluRS的基因互补。重组Arc1p在体外与纯化的单体酵母MetRS结合，但不与N端截短形式结合，并强烈增加其对tRNA Met的表观亲和力。此外，与四链体核酸结合蛋白G4p1等位的Arc1p，通过凝胶阻滞和紫外线交联测定，显示出对tRNA的特异性结合。因此，Arc1p是一种具有双重特异性的酵母蛋白：它与tRNA和氨酰-tRNA合成酶相关联。这种功能相互作用可能是体内有效氨酰化所必需的。
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