Related Experiment Video
Updated: May 30, 2026
07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
A new basal promoter element recognized by RNA polymerase core enzyme
Yulia Yuzenkova1, Vasisht R Tadigotla, Konstantin Severinov
1Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, UK.
The EMBO Journal
|July 28, 2011
Summary
The bacterial RNA polymerase β
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Bacterial RNA polymerase (RNAP) holoenzyme, specifically the sigma (σ) subunit, recognizes promoter elements (-10 and -35 regions) to initiate transcription.
- The precise mechanisms governing promoter recognition and the formation of stable transcription-initiation complexes are complex and involve multiple RNAP subunits.
- The role of conserved structural elements within the RNAP core enzyme, beyond the σ subunit, in promoter interaction remains an area of active investigation.
Purpose of the Study:
- To investigate the role of the β' zipper, a conserved loop in the largest RNAP subunit, in bacterial promoter recognition and transcription initiation.
- To determine if the β' zipper interacts with the promoter spacer DNA and influences the formation of closed and open promoter complexes.
- To explore the potential for β' zipper-DNA interactions to contribute to promoter activity and transcription regulation, including RNAP pausing.
Main Methods:
- Utilized biochemical assays to probe the interaction between the RNAP β' zipper and promoter DNA sequences.
- Investigated the effect of β' zipper-promoter spacer interactions on the formation of closed and open promoter complexes.
- Analyzed the impact of these interactions on transcription initiation and promoter-proximal RNAP pausing.
Main Results:
- Provided evidence for direct interaction between the bacterial RNAP β' zipper and the promoter spacer DNA.
- Demonstrated that this interaction facilitates the formation of stable closed promoter complexes.
- Showed that β' zipper-spacer interactions can influence open promoter complex formation and, in some cases, substitute for σ subunit interactions with the -35 element, suggesting a novel class of promoters.
Conclusions:
- The β' zipper is a key mediator of bacterial promoter recognition, interacting with the promoter spacer DNA.
- This interaction contributes to the stability of transcription initiation complexes and can modulate promoter activity.
- Sequence-dependent β' zipper-DNA interactions play a role in regulating transcription through promoter-proximal RNAP pausing.
Related Concept Videos
02:40The Eukaryotic Promoter Region
The eukaryotic promoter region is a segment of DNA located upstream of a gene. It contains an RNA polymerase binding site, a transcription start site, and several cis-regulatory sequences. The proximal promoter region is located in the vicinity of the gene and has cis-regulatory sequences and the core promoter. The core promoter is the binding site for RNA polymerase and is usually located between -35 and +35 nucleotides from the transcription start site. The distal promoter regions are...
02:40The Eukaryotic Promoter Region
The eukaryotic promoter region is a segment of DNA located upstream of a gene. It contains an RNA polymerase binding site, a transcription start site, and several cis-regulatory sequences. The proximal promoter region is located in the vicinity of the gene and has cis-regulatory sequences and the core promoter. The core promoter is the binding site for RNA polymerase and is usually located between -35 and +35 nucleotides from the transcription start site. The distal promoter regions are...
01:47Transcription Initiation
Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.
The promoters and enhancers and their accessory proteins allow tight regulation of...
The promoters and enhancers and their accessory proteins allow tight regulation of...
00:43Bacterial 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...
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...
00:43Bacterial 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...
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...
01:53Bacterial 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:
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are: