Kinetic Analysis Suggests Evolution of Ribosome Specificity in Modern Elongation Factor-Tus from 'Generalist' Ancestors
| Autor: | Arindam, De Tarafder, Narayan Prasad, Parajuli, Soneya, Majumdar, Betül, Kaçar, Suparna, Sanyal |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
molecular evolution
Thermus thermophilus AcademicSubjects/SCI01130 Peptide Elongation Factor Tu AcademicSubjects/SCI01180 Substrate Specificity Evolution Molecular Kinetics Bacterial Proteins Protein Biosynthesis fast kinetics specificity Escherichia coli ancestral sequence reconstruction generalist Ribosomes Discoveries translation machinery EF-Tu |
| Zdroj: | Molecular Biology and Evolution |
| ISSN: | 1537-1719 |
| Popis: | It has been hypothesized that early enzymes are more promiscuous than their extant orthologs. Whether or not this hypothesis applies to the translation machinery, the oldest molecular machine of life, is not known. Efficient protein synthesis relies on a cascade of specific interactions between the ribosome and the translation factors. Here, using elongation factor-Tu (EF-Tu) as a model system, we have explored the evolution of ribosome specificity in translation factors. Employing presteady state fast kinetics using quench flow, we have quantitatively characterized the specificity of two sequence-reconstructed 1.3- to 3.3-Gy-old ancestral EF-Tus toward two unrelated bacterial ribosomes, mesophilic Escherichia coli and thermophilic Thermus thermophilus. Although the modern EF-Tus show clear preference for their respective ribosomes, the ancestral EF-Tus show similar specificity for diverse ribosomes. In addition, despite increase in the catalytic activity with temperature, the ribosome specificity of the thermophilic EF-Tus remains virtually unchanged. Our kinetic analysis thus suggests that EF-Tu proteins likely evolved from the catalytically promiscuous, “generalist” ancestors. Furthermore, compatibility of diverse ribosomes with the modern and ancestral EF-Tus suggests that the ribosomal core probably evolved before the diversification of the EF-Tus. This study thus provides important insights regarding the evolution of modern translation machinery. |
| Databáze: | OpenAIRE |
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