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Autonomous synthesis and assembly of a ribosomal subunit on a chip.

ABSTRACT: Ribosome biogenesis is an efficient and complex assembly process that has not been reconstructed outside a living cell so far, yet is the most critical step for establishing a self-replicating artificial cell. We recreated the biogenesis of Escherichia coli's small ribosomal subunit by synthesizing and capturing all its ribosomal proteins and RNA on a chip. Surface confinement provided favorable conditions for autonomous stepwise assembly of new subunits, spatially segregated from original intact ribosomes. Our real-time fluorescence measurements revealed hierarchal assembly, cooperative interactions, unstable intermediates, and specific binding to large ribosomal subunits. Using only synthetic genes, our methodology is a crucial step toward creation of a self-replicating artificial cell and a general strategy for the mechanistic investigation of diverse multicomponent macromolecular machines.

SUBMITTER: Levy M

PROVIDER: S-EPMC7159907 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Autonomous synthesis and assembly of a ribosomal subunit on a chip.

Levy Michael M Falkovich Reuven R Daube Shirley S SS Bar-Ziv Roy H RH

Science advances 20200415 16

Ribosome biogenesis is an efficient and complex assembly process that has not been reconstructed outside a living cell so far, yet is the most critical step for establishing a self-replicating artificial cell. We recreated the biogenesis of <i>Escherichia coli's</i> small ribosomal subunit by synthesizing and capturing all its ribosomal proteins and RNA on a chip. Surface confinement provided favorable conditions for autonomous stepwise assembly of new subunits, spatially segregated from origina ...[more]

PMID: 32494616

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