A transcriptional activator is a protein (transcription factor) that increases transcription of a gene or set of genes. [1] Activators are considered to have positive control over gene expression, as they function to promote gene transcription and, in some cases, are required for the transcription of genes to occur. [1] [2] [3] [4] Most activators are DNA-binding proteins that bind to enhancers or promoter-proximal elements. [1] The DNA site bound by the activator is referred to as an "activator-binding site". [3] The part of the activator that makes protein–protein interactions with the general transcription machinery is referred to as an "activating region" or "activation domain". [1]
Most activators function by binding sequence-specifically to a regulatory DNA site located near a promoter and making protein–protein interactions with the general transcription machinery (RNA polymerase and general transcription factors), thereby facilitating the binding of the general transcription machinery to the promoter. [1] [2] [3] [4] Other activators help promote gene transcription by triggering RNA polymerase to release from the promoter and proceed along the DNA. [2] At times, RNA polymerase can pause shortly after leaving the promoter; activators also function to allow these “stalled” RNA polymerases to continue transcription. [1] [2]
The activity of activators can be regulated. Some activators have an allosteric site and can only function when a certain molecule binds to this site, essentially turning the activator on. [4] Post-translational modifications to activators can also regulate activity, increasing or decreasing activity depending on the type of modification and activator being modified. [1]
In some cells, usually eukaryotes, multiple activators can bind to the binding-site; these activators tend to bind cooperatively and interact synergistically. [1] [2]
Activator proteins consist of two main domains: a DNA-binding domain that binds to a DNA sequence specific to the activator, and an activation domain that functions to increase gene transcription by interacting with other molecules. [1] Activator DNA-binding domains come in a variety of conformations, including the helix-turn-helix, zinc finger, and leucine zipper among others. [1] [2] [3] These DNA-binding domains are specific to a certain DNA sequence, allowing activators to turn on only certain genes. [1] [2] [3] Activation domains also come in a variety of types that are categorized based on the domain's amino acid sequence, including alanine-rich, glutamine-rich, and acidic domains. [1] These domains are not as specific, and tend to interact with a variety of target molecules. [1]
Activators can also have allosteric sites that are responsible for turning the activators themselves on and off. [4]
Within the grooves of the DNA double helix, functional groups of the base pairs are exposed. [2] The sequence of the DNA thus creates a unique pattern of surface features, including areas of possible hydrogen bonding, ionic bonding, as well as hydrophobic interactions. [2] Activators also have unique sequences of amino acids with side chains that are able to interact with the functional groups in DNA. [2] [3] Thus, the pattern of amino acid side chains making up an activator protein will be complementary to the surface features of the specific DNA regulatory sequence it was designed to bind to. [1] [2] [3] The complementary interactions between the amino acids of the activator protein and the functional groups of the DNA create an “exact-fit” specificity between the activator and its regulatory DNA sequence. [2]
Most activators bind to the major grooves of the double helix, as these areas tend to be wider, but there are some that will bind to the minor grooves. [1] [2] [3]
Activator-binding sites may be located very close to the promoter or numerous base pairs away. [2] [3] If the regulatory sequence is located far away, the DNA will loop over itself (DNA looping) in order for the bound activator to interact with the transcription machinery at the promoter site. [2] [3]
In prokaryotes, multiple genes can be transcribed together (operon), and are thus controlled under the same regulatory sequence. [2] In eukaryotes, genes tend to be transcribed individually, and each gene is controlled by its own regulatory sequences. [2] Regulatory sequences where activators bind are commonly found upstream from the promoter, but they can also be found downstream or even within introns in eukaryotes. [1] [2] [3]
Binding of the activator to its regulatory sequence promotes gene transcription by enabling RNA polymerase activity. [1] [2] [3] [4] This is done through various mechanisms, such as recruiting transcription machinery to the promoter and triggering RNA polymerase to continue into elongation. [1] [2] [3] [4]
Activator-controlled genes require the binding of activators to regulatory sites in order to recruit the necessary transcription machinery to the promoter region. [1] [2] [3]
Activator interactions with RNA polymerase are mostly direct in prokaryotes and indirect in eukaryotes. [2] In prokaryotes, activators tend to make contact with the RNA polymerase directly in order to help bind it to the promoter. [2] In eukaryotes, activators mostly interact with other proteins, and these proteins will then be the ones to interact with the RNA polymerase. [2]
In prokaryotes, genes controlled by activators have promoters that are unable to strongly bind to RNA polymerase by themselves. [2] [3] Thus, activator proteins help to promote the binding of the RNA polymerase to the promoter. [2] [3] This is done through various mechanisms. Activators may bend the DNA in order to better expose the promoter so the RNA polymerase can bind more effectively. [3] Activators may make direct contact with the RNA polymerase and secure it to the promoter. [2] [3] [4]
In eukaryotes, activators have a variety of different target molecules that they can recruit in order to promote gene transcription. [1] [2] They can recruit other transcription factors and cofactors that are needed in transcription initiation. [1] [2]
Activators can recruit molecules known as coactivators. [1] [2] These coactivator molecules can then perform functions necessary for beginning transcription in place of the activators themselves, such as chromatin modifications. [1] [2]
DNA is much more condensed in eukaryotes; thus, activators tend to recruit proteins that are able to restructure the chromatin so the promoter is more easily accessible by the transcription machinery. [1] [2] Some proteins will rearrange the layout of nucleosomes along the DNA in order to expose the promoter site (ATP-dependent chromatin remodeling complexes). [1] [2] Other proteins affect the binding between histones and DNA via post-translational histone modifications, allowing the DNA tightly wrapped into nucleosomes to loosen. [1] [2]
All of these recruited molecules work together in order to ultimately recruit the RNA polymerase to the promoter site. [1] [2]
Activators can promote gene transcription by signaling the RNA polymerase to move beyond the promoter and proceed along the DNA, initiating the beginning of transcription. [2] The RNA polymerase can sometimes pause shortly after beginning transcription, and activators are required to release RNA polymerase from this “stalled” state. [1] [2] Multiple mechanisms exist for releasing these “stalled” RNA polymerases. Activators may act simply as a signal to trigger the continued movement of the RNA polymerase. [2] If the DNA is too condensed to allow RNA polymerase to continue transcription, activators may recruit proteins that can restructure the DNA so any blocks are removed. [1] [2] Activators may also promote the recruitment of elongation factors, which are necessary for the RNA polymerase to continue transcription. [1] [2]
There are different ways in which the activity of activators themselves can be regulated, in order to ensure that activators are stimulating gene transcription at appropriate times and levels. [1] Activator activity can increase or decrease in response to environmental stimuli or other intracellular signals. [1]
Activators often must be “turned on” before they can promote gene transcription. [2] [3] [4] The activity of activators is controlled by the ability of the activator to bind to its regulatory site along the DNA. [2] [3] [4] The DNA-binding domain of the activator has an active form and an inactive form, which are controlled by the binding of molecules known as allosteric effectors to the allosteric site of the activator. [4]
Activators in their inactive form are not bound to any allosteric effectors. [4] When inactive, the activator is unable to bind to its specific regulatory sequence in the DNA, and thus has no regulatory effect on the transcription of genes. [4]
When an allosteric effector binds to the allosteric site of an activator, a conformational change in the DNA-binding domain occurs, which allows the protein to bind to the DNA and increase gene transcription. [2] [4]
Some activators are able to undergo post-translational modifications that have an effect on their activity within a cell. [1] Processes such as phosphorylation, acetylation, and ubiquitination, among others, have been seen to regulate the activity of activators. [1] Depending on the chemical group being added, as well as the nature of the activator itself, post-translational modifications can either increase or decrease the activity of an activator. [1] For example, acetylation has been seen to increase the activity of some activators through mechanisms such as increasing DNA-binding affinity. [1] On the other hand, ubiquitination decreases the activity of activators, as ubiquitin marks proteins for degradation after they have performed their respective functions. [1]
In prokaryotes, a lone activator protein is able to promote transcription. [2] [3] In eukaryotes, usually more than one activator assembles at the binding-site, forming a complex that acts to promote transcription. [1] [2] These activators bind cooperatively at the binding-site, meaning that the binding of one activator increases the affinity of the site to bind another activator (or in some cases another transcriptional regulator) thus making it easier for multiple activators to bind at the site. [1] [2] In these cases, the activators interact with each other synergistically, meaning that the rate of transcription that is achieved from multiple activators working together is much higher than the additive effects of the activators if they were working individually. [1] [2]
The breakdown of maltose in Escherichia coli is controlled by gene activation. [3] The genes that code for the enzymes responsible for maltose catabolism can only be transcribed in the presence of an activator. [3] The activator that controls transcription of the maltose enzymes is “off” in the absence of maltose. [3] In its inactive form, the activator is unable to bind to DNA and promote transcription of the maltose genes. [3] [4]
When maltose is present in the cell, it binds to the allosteric site of the activator protein, causing a conformational change in the DNA-binding domain of the activator. [3] [4] This conformational change “turns on” the activator by allowing it to bind to its specific regulatory DNA sequence. [3] [4] Binding of the activator to its regulatory site promotes RNA polymerase binding to the promoter and thus transcription, producing the enzymes that are needed to break down the maltose that has entered the cell. [3]
The catabolite activator protein (CAP), otherwise known as cAMP receptor protein (CRP), activates transcription at the lac operon of the bacterium Escherichia coli. [5] Cyclic adenosine monophosphate (cAMP) is produced during glucose starvation; this molecule acts as an allosteric effector that binds to CAP and causes a conformational change that allows CAP to bind to a DNA site located adjacent to the lac promoter. [5] CAP then makes a direct protein–protein interaction with RNA polymerase that recruits RNA polymerase to the lac promoter. [5]
In genetics, a promoter is a sequence of DNA to which proteins bind to initiate transcription of a single RNA transcript from the DNA downstream of the promoter. The RNA transcript may encode a protein (mRNA), or can have a function in and of itself, such as tRNA or rRNA. Promoters are located near the transcription start sites of genes, upstream on the DNA . Promoters can be about 100–1000 base pairs long, the sequence of which is highly dependent on the gene and product of transcription, type or class of RNA polymerase recruited to the site, and species of organism.
Gene expression is the process by which information from a gene is used in the synthesis of a functional gene product that enables it to produce end products, protein or non-coding RNA, and ultimately affect a phenotype, as the final effect. These products are often proteins, but in non-protein-coding genes such as transfer RNA (tRNA) and small nuclear RNA (snRNA), the product is a functional non-coding RNA. Gene expression is summarized in the central dogma of molecular biology first formulated by Francis Crick in 1958, further developed in his 1970 article, and expanded by the subsequent discoveries of reverse transcription and RNA replication.
Transcription is the process of copying a segment of DNA into RNA. The segments of DNA transcribed into RNA molecules that can encode proteins are said to produce messenger RNA (mRNA). Other segments of DNA are copied into RNA molecules called non-coding RNAs (ncRNAs). mRNA comprises only 1–3% of total RNA samples. Less than 2% of the human genome can be transcribed into mRNA, while at least 80% of mammalian genomic DNA can be actively transcribed, with the majority of this 80% considered to be ncRNA.
In genetics, an operon is a functioning unit of DNA containing a cluster of genes under the control of a single promoter. The genes are transcribed together into an mRNA strand and either translated together in the cytoplasm, or undergo splicing to create monocistronic mRNAs that are translated separately, i.e. several strands of mRNA that each encode a single gene product. The result of this is that the genes contained in the operon are either expressed together or not at all. Several genes must be co-transcribed to define an operon.
A regulatory sequence is a segment of a nucleic acid molecule which is capable of increasing or decreasing the expression of specific genes within an organism. Regulation of gene expression is an essential feature of all living organisms and viruses.
In molecular biology and genetics, transcriptional regulation is the means by which a cell regulates the conversion of DNA to RNA (transcription), thereby orchestrating gene activity. A single gene can be regulated in a range of ways, from altering the number of copies of RNA that are transcribed, to the temporal control of when the gene is transcribed. This control allows the cell or organism to respond to a variety of intra- and extracellular signals and thus mount a response. Some examples of this include producing the mRNA that encode enzymes to adapt to a change in a food source, producing the gene products involved in cell cycle specific activities, and producing the gene products responsible for cellular differentiation in multicellular eukaryotes, as studied in evolutionary developmental biology.
In molecular biology, the TATA box is a sequence of DNA found in the core promoter region of genes in archaea and eukaryotes. The bacterial homolog of the TATA box is called the Pribnow box which has a shorter consensus sequence.
Regulation of gene expression, or gene regulation, includes a wide range of mechanisms that are used by cells to increase or decrease the production of specific gene products. Sophisticated programs of gene expression are widely observed in biology, for example to trigger developmental pathways, respond to environmental stimuli, or adapt to new food sources. Virtually any step of gene expression can be modulated, from transcriptional initiation, to RNA processing, and to the post-translational modification of a protein. Often, one gene regulator controls another, and so on, in a gene regulatory network.
In molecular genetics, a repressor is a DNA- or RNA-binding protein that inhibits the expression of one or more genes by binding to the operator or associated silencers. A DNA-binding repressor blocks the attachment of RNA polymerase to the promoter, thus preventing transcription of the genes into messenger RNA. An RNA-binding repressor binds to the mRNA and prevents translation of the mRNA into protein. This blocking or reducing of expression is called repression.
In genetics, a silencer is a DNA sequence capable of binding transcription regulation factors, called repressors. DNA contains genes and provides the template to produce messenger RNA (mRNA). That mRNA is then translated into proteins. When a repressor protein binds to the silencer region of DNA, RNA polymerase is prevented from transcribing the DNA sequence into RNA. With transcription blocked, the translation of RNA into proteins is impossible. Thus, silencers prevent genes from being expressed as proteins.
A regulator gene, regulator, or regulatory gene is a gene involved in controlling the expression of one or more other genes. Regulatory sequences, which encode regulatory genes, are often at the five prime end (5') to the start site of transcription of the gene they regulate. In addition, these sequences can also be found at the three prime end (3') to the transcription start site. In both cases, whether the regulatory sequence occurs before (5') or after (3') the gene it regulates, the sequence is often many kilobases away from the transcription start site. A regulator gene may encode a protein, or it may work at the level of RNA, as in the case of genes encoding microRNAs. An example of a regulator gene is a gene that codes for a repressor protein that inhibits the activity of an operator.
In molecular biology, an inducer is a molecule that regulates gene expression. An inducer functions in two ways; namely:
A coactivator is a type of transcriptional coregulator that binds to an activator to increase the rate of transcription of a gene or set of genes. The activator contains a DNA binding domain that binds either to a DNA promoter site or a specific DNA regulatory sequence called an enhancer. Binding of the activator-coactivator complex increases the speed of transcription by recruiting general transcription machinery to the promoter, therefore increasing gene expression. The use of activators and coactivators allows for highly specific expression of certain genes depending on cell type and developmental stage.
Gene structure is the organisation of specialised sequence elements within a gene. Genes contain most of the information necessary for living cells to survive and reproduce. In most organisms, genes are made of DNA, where the particular DNA sequence determines the function of the gene. A gene is transcribed (copied) from DNA into RNA, which can either be non-coding (ncRNA) with a direct function, or an intermediate messenger (mRNA) that is then translated into protein. Each of these steps is controlled by specific sequence elements, or regions, within the gene. Every gene, therefore, requires multiple sequence elements to be functional. This includes the sequence that actually encodes the functional protein or ncRNA, as well as multiple regulatory sequence regions. These regions may be as short as a few base pairs, up to many thousands of base pairs long.
The L-arabinose operon, also called the ara or araBAD operon, is an operon required for the breakdown of the five-carbon sugar L-arabinose in Escherichia coli. The L-arabinose operon contains three structural genes: araB, araA, araD, which encode for three metabolic enzymes that are required for the metabolism of L-arabinose. AraB (ribulokinase), AraA, AraD produced by these genes catalyse conversion of L-arabinose to an intermediate of the pentose phosphate pathway, D-xylulose-5-phosphate.
cAMP receptor protein is a regulatory protein in bacteria. CRP protein binds cAMP, which causes a conformational change that allows CRP to bind tightly to a specific DNA site in the promoters of the genes it controls. CRP then activates transcription through direct protein–protein interactions with RNA polymerase.
Eukaryotic transcription is the elaborate process that eukaryotic cells use to copy genetic information stored in DNA into units of transportable complementary RNA replica. Gene transcription occurs in both eukaryotic and prokaryotic cells. Unlike prokaryotic RNA polymerase that initiates the transcription of all different types of RNA, RNA polymerase in eukaryotes comes in three variations, each translating a different type of gene. A eukaryotic cell has a nucleus that separates the processes of transcription and translation. Eukaryotic transcription occurs within the nucleus where DNA is packaged into nucleosomes and higher order chromatin structures. The complexity of the eukaryotic genome necessitates a great variety and complexity of gene expression control.
RNA polymerase II holoenzyme is a form of eukaryotic RNA polymerase II that is recruited to the promoters of protein-coding genes in living cells. It consists of RNA polymerase II, a subset of general transcription factors, and regulatory proteins known as SRB proteins.
The 5′ flanking region is a region of DNA that is adjacent to the 5′ end of the gene. The 5′ flanking region contains the promoter, and may contain enhancers or other protein binding sites. It is the region of DNA that is not transcribed into RNA. Not to be confused with the 5′ untranslated region, this region is not transcribed into RNA or translated into a functional protein. These regions primarily function in the regulation of gene transcription. 5′ flanking regions are categorized between prokaryotes and eukaryotes.
Promoter activity is a term that encompasses several meanings around the process of gene expression from regulatory sequences —promoters and enhancers. Gene expression has been commonly characterized as a measure of how much, how fast, when and where this process happens. Promoters and enhancers are required for controlling where and when a specific gene is transcribed.