Real-time assembly of ribonucleoprotein complexes on nascent RNA transcripts.

Abstract:

:Cellular protein-RNA complexes assemble on nascent transcripts, but methods to observe transcription and protein binding in real time and at physiological concentrations are not available. Here, we report a single-molecule approach based on zero-mode waveguides that simultaneously tracks transcription progress and the binding of ribosomal protein S15 to nascent RNA transcripts during early ribosome biogenesis. We observe stable binding of S15 to single RNAs immediately after transcription for the majority of the transcripts at 35 °C but for less than half at 20 °C. The remaining transcripts exhibit either rapid and transient binding or are unable to bind S15, likely due to RNA misfolding. Our work establishes the foundation for studying transcription and its coupled co-transcriptional processes, including RNA folding, ligand binding, and enzymatic activity such as in coupling of transcription to splicing, ribosome assembly or translation.

journal_name

Nat Commun

journal_title

Nature communications

authors

Duss O,Stepanyuk GA,Grot A,O'Leary SE,Puglisi JD,Williamson JR

doi

10.1038/s41467-018-07423-3

subject

Has Abstract

pub_date

2018-11-30 00:00:00

pages

5087

issue

1

issn

2041-1723

pii

10.1038/s41467-018-07423-3

journal_volume

9

pub_type

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