Real-time assembly of ribonucleoprotein complexes on nascent RNA transcripts
| Autor: | Sean T. O’Leary, Galina A. Stepanyuk, Olivier Duss, James R. Williamson, Annette Grot, Joseph D. Puglisi |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Ribosomal Proteins RNA Folding Science RNA Splicing 1.1 Normal biological development and functioning General Physics and Astronomy Ribosome biogenesis Plasma protein binding General Biochemistry Genetics and Molecular Biology Article Ribosome assembly 03 medical and health sciences 0302 clinical medicine Transcription (biology) Ribosomal protein Underpinning research Genetics lcsh:Science Ribonucleoprotein Ribosomal Multidisciplinary Chemistry RNA General Chemistry 3. Good health Cell biology 030104 developmental biology RNA Ribosomal RNA splicing Nucleic Acid Conformation lcsh:Q Generic health relevance 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | Nature communications, vol 9, iss 1 Nature Communications Nature Communications, Vol 9, Iss 1, Pp 1-14 (2018) |
| Popis: | 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. The early steps of ribosome assembly occur co-transcriptionally on the nascent ribosomal RNA. Here the authors demonstrate an approach that allows simultaneous monitoring of transcription and ribosomal protein assembly at the single-molecule level in real time. |
| Databáze: | OpenAIRE |
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