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Bacterial RNA Polymerase

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...

Bacterial RNA Polymerase

Bacterial RNA Polymerase

Bacterial Transcription

Bacterial Transcription

RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:

RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...

Ribozymes

Ribozymes

The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can...

Eukaryotic RNA Polymerases

Eukaryotic RNA Polymerases

RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...

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Related Experiment Video

Updated: Mar 7, 2026

Chemical Triphosphorylation of Oligonucleotides

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

Evolution of a split RNA polymerase as a versatile biosensor platform.

Jinyue Pu¹, Julia Zinkus-Boltz¹, Bryan C Dickinson¹

¹Department of Chemistry, The University of Chicago, Chicago, Illinois, USA.

Nature Chemical Biology

|February 14, 2017

Summary

This summary is machine-generated.

Researchers developed a novel split RNA polymerase (RNAP) biosensor platform. This system efficiently detects inputs and performs synthetic biology functions in mammalian cells.

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Area of Science:

Synthetic Biology
Bioengineering
Molecular Biology

Background:

Biosensors are crucial for bioengineering, converting chemical/biochemical signals into genetic outputs.
Existing biosensors face challenges like low signal-to-noise, extensive optimization, and poor mammalian cell performance.

Purpose of the Study:

To develop a general platform for biosensor engineering using a proximity-dependent split RNA polymerase (RNAP).
To overcome limitations of current biosensor designs, enhancing signal detection and performance in mammalian systems.

Main Methods:

Engineered a split T7 RNA polymerase (RNAP) system responsive to protein-protein interactions.
Utilized phage-assisted continuous evolution (PACE) to optimize split RNAP components for interaction detection.
Developed activity-responsive RNAP (AR) systems for biosensor applications.

Main Results:

Demonstrated a 'plug-and-play' biosensor platform activated by light and small molecules.
Validated AR system's ability to detect multidimensional protein-protein interactions.
Showcased ARs triggering RNA nanostructures, protein synthesis, and gene knockdown in mammalian cells.

Conclusions:

The proximity-dependent split RNAP platform offers a versatile and efficient tool for biosensor engineering.
The activity-responsive RNAP (AR) system significantly advances synthetic biology applications in mammalian systems.
This technology provides a robust foundation for designing next-generation biosensors with improved functionality.