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The Eukaryotic Promoter Region

The Eukaryotic Promoter Region

The Eukaryotic Promoter Region

The 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...

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 Polymerase II Accessory Proteins

RNA Polymerase II Accessory Proteins

Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...

Transcription Attenuation in Prokaryotes

Transcription Attenuation in Prokaryotes

Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure. Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...

Transcription Initiation

Transcription 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...

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Related Experiment Video

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Rapid Verification of Terminators Using the pGR-Blue Plasmid and Golden Gate Assembly

Rapid Verification of Terminators Using the pGR-Blue Plasmid and Golden Gate Assembly

Published on: April 25, 2016

Promoter and Terminator Discovery and Engineering.

Matthew Deaner¹, Hal S Alper^2,3

¹McKetta Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton St. Stop C0400, Austin, TX, 78712, USA.

Advances in Biochemical Engineering/Biotechnology

|June 10, 2016

Summary

Discovering and engineering DNA elements like promoters and terminators is key for synthetic biology. New methods enable precise control over gene expression in various organisms.

Keywords:

Biotechnology Gene expression Metabolic engineering Promoters Synthetic biology Terminators

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Area of Science:

Synthetic Biology
Molecular Biology
Biotechnology

Background:

Gene expression control is vital for optimizing metabolic pathways and synthetic gene networks.
Promoters and terminators are DNA elements regulating transcription rates and mRNA degradation, thus controlling protein expression.

Purpose of the Study:

To highlight diverse approaches for cataloguing synthetic gene regulatory elements.
To review methodologies for discovering and engineering promoters and terminators with desired characteristics.

Main Methods:

Semi-rational techniques like saturation mutagenesis for diversifying native elements.
Rational design strategies including hybrid engineering and thermodynamic modeling.
High-throughput characterization for efficient screening and design.

Main Results:

Cataloguing of synthetic promoters and terminators has evolved from semi-rational to rational design approaches.
Advanced methods allow for finer control in designing novel promoters and terminators.
Demonstrated utility of these methods across multiple host organisms.

Conclusions:

Engineering synthetic promoters and terminators is crucial for advancing synthetic biology.
Rational design and high-throughput methods accelerate the discovery and optimization of gene regulatory elements.
These engineered elements offer precise control over gene expression in diverse biological systems.