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Capturing RNA-dependent pathways for cryo-EM analysis.

ABSTRACT: Cryo-Electron Microscopy (EM) is a powerful technique to visualize biological processes at nanometer resolution. Structural studies of macromolecular assemblies are typically performed on individual complexes that are biochemically isolated from their cellular context. Here we present a molecular imaging platform to capture and view multiple components of cellular pathways within a functionally relevant framework. We utilized the bacterial protein synthesis machinery as a model system to develop our approach. By using modified Affinity Grid surfaces, we were able to recruit multiple protein assemblies bound to nascent strands of mRNA. The combined use of Affinity Capture technology and single particle electron microscopy provide the basis for visualizing RNA-dependent pathways in a remarkable new way.

SUBMITTER: Tanner JR

PROVIDER: S-EPMC3962177 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Capturing RNA-dependent pathways for cryo-EM analysis.

Tanner Justin R JR Degen Katherine K Gilmore Brian L BL Kelly Deborah F DF

Computational and structural biotechnology journal 20120223

Cryo-Electron Microscopy (EM) is a powerful technique to visualize biological processes at nanometer resolution. Structural studies of macromolecular assemblies are typically performed on individual complexes that are biochemically isolated from their cellular context. Here we present a molecular imaging platform to capture and view multiple components of cellular pathways within a functionally relevant framework. We utilized the bacterial protein synthesis machinery as a model system to develop ...[more]

PMID: 24688633

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