Co‐transcriptional assembly mechanisms of <scp>protein‐RNA</scp> complexes

Qureshi, Nusrat S.; Duss, Olivier

doi:10.1002/1873-3468.14758

Cited by 4 publications

(1 citation statement)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Co-transcriptional RNA looping is likely a more general feature for increasing the efficiency of transcription-coupled processes 46 . Both in bacterial rRNA transcription (mediated by interactions between the ribosomal RNA transcription antitermination complex and the RNAP 47 ) and eukaryotic co-transcriptional splicing (mediated by a U1 snRNP – Pol-II interactions 48 ) transcription-mediated RNA looping could provide means for increased RNA processing efficiency (RNase III processing and splicing, respectively).…”

Section: Discussionmentioning

confidence: 99%

Tracking transcription-translation coupling in real-time

Qureshi,

Duss

2023

Preprint

Self Cite

View full text Add to dashboard Cite

A central question in biology is how macromolecular machines function cooperatively. In bacteria, transcription and translation occur in the same cellular compartment and can be physically and functionally coupled. While several recently published high-resolution structures of the ribosome-RNA polymerase (RNAP) complex provided first mechanistic insight into the coupling process, we do not know how these structural snapshots are placed along a dynamic reaction trajectory. Here, we reconstitute a complete active transcription-translation system and develop multi-color single-molecule fluorescence microscopy experiments to directly and simultaneously track transcription elongation, translation elongation and the physical and functional coupling between the ribosome and the RNAP in real-time. Our data show that the ribosome slows down while colliding with the RNAP and that coupling following a collision becomes less efficient. Unexpectedly, physical coupling can occur with hundreds of nucleotides of intervening mRNA between both machineries by mRNA looping, and increases in efficiency in presence of NusG. We detect active transcription elongation during mRNA looping and show that NusA-paused RNAPs can be activated by the ribosome by long-range physical coupling. We provide an alternative explanation on how the ribosome can rescue RNAP from frequent pausing without requiring collisions by a closely trailing ribosome. Our data mechanistically highlight an example of how macromolecular machines central to gene expression physically and functionally cooperate.

show abstract

Section: Discussionmentioning

confidence: 99%

Tracking transcription-translation coupling in real-time

Qureshi,

Duss

2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Beyond ligand binding: Single molecule observation reveals how riboswitches integrate multiple signals to balance bacterial gene regulation

Chauvier,

Walter

2024

Current Opinion in Structural Biology

View full text Add to dashboard Cite

Tracking transcription–translation coupling in real time

Qureshi,

Duss

2024

Nature

View full text Add to dashboard Cite

A central question in biology is how macromolecular machines function cooperatively. In bacteria, transcription and translation occur in the same cellular compartment, and can be physically and functionally coupled1–4. Although high-resolution structures of the ribosome–RNA polymerase (RNAP) complex have provided initial mechanistic insights into the coupling process5–10, we lack knowledge of how these structural snapshots are placed along a dynamic reaction trajectory. Here we reconstitute a complete and active transcription–translation system and develop multi-colour single-molecule fluorescence microscopy experiments to directly and simultaneously track transcription elongation, translation elongation and the physical and functional coupling between the ribosome and the RNAP in real time. Our data show that physical coupling between ribosome and RNAP can occur over hundreds of nucleotides of intervening mRNA by mRNA looping, a process facilitated by NusG. We detect active transcription elongation during mRNA looping and show that NusA-paused RNAPs can be activated by the ribosome by long-range physical coupling. Conversely, the ribosome slows down while colliding with the RNAP. We hereby provide an alternative explanation for how the ribosome can efficiently rescue RNAP from frequent pausing without requiring collisions by a closely trailing ribosome. Overall, our dynamic data mechanistically highlight an example of how two central macromolecular machineries, the ribosome and RNAP, can physically and functionally cooperate to optimize gene expression.

show abstract

Co‐transcriptional assembly mechanisms of protein‐RNA complexes

Cited by 4 publications

References 29 publications

Tracking transcription-translation coupling in real-time

Tracking transcription-translation coupling in real-time

Beyond ligand binding: Single molecule observation reveals how riboswitches integrate multiple signals to balance bacterial gene regulation

Tracking transcription–translation coupling in real time

Contact Info

Product

Resources

About