Autonomous biogenesis of the entire protein translation machinery excluding ribosomes
Recreating the conditions for autonomous biogenesis of the protein translation machinery is fundamental to our understanding of living systems and is essential for the assembly of minimal cell models. All components of the translation machinery, including the ribosomes, translation factors and aminoacyl tRNA synthetases, are made of proteins and are therefore involved in their own synthesis, posing a unique challenge for self-biogenesis. Here, we created physicochemical conditions for autonomous biogenesis of all the translation machinery, excluding the ribosome. We surface-immobilized synthetic genes coding for all thirty components as densely packed DNA brushes forming a reaction center that localizes, concentrates and catalyzes their simultaneous synthesis. To demonstrate their activity, we first determined empirically the minimal critical concentration of each translation protein required to initiate its own self-biogenesis in bulk solution. We then assembled a minimal gene expression reaction with all translation proteins at their critical concentrations. Under these dilute conditions, reporter proteins were not synthesized unless the DNA brushes encoded all the translation proteins, thereby demonstrating their co-synthesis, functionality and engagement in their own synthesis. This scenario of a complex biochemical process that amplifies itself can be generalized and extended to impact our understanding toward the design of autonomous self-replicating biological system.