Autoinducer

Autoinducers are signaling molecules that are produced in response to changes in cell-population density. As the density of quorum sensing bacterial cells increases so does the concentration of the autoinducer. Detection of signal molecules by bacteria acts as stimulation which leads to altered gene expression once the minimal threshold is reached. Quorum sensing is a phenomenon that allows both Gram-negative and Gram-positive bacteria to sense one another and to regulate a wide variety of physiological activities. Such activities include symbiosis, virulence, motility, antibiotic production, and biofilm formation. Autoinducers come in a number of different forms depending on the species, but the effect that they have is similar in many cases. Autoinducers allow bacteria to communicate both within and between different species. This communication alters gene expression and allows bacteria to mount coordinated responses to their environments, in a manner that is comparable to behavior and signaling in higher organisms. Not surprisingly, it has been suggested that quorum sensing may have been an important evolutionary milestone that ultimately gave rise to multicellular life forms. Autoinducers are signaling molecules that are produced in response to changes in cell-population density. As the density of quorum sensing bacterial cells increases so does the concentration of the autoinducer. Detection of signal molecules by bacteria acts as stimulation which leads to altered gene expression once the minimal threshold is reached. Quorum sensing is a phenomenon that allows both Gram-negative and Gram-positive bacteria to sense one another and to regulate a wide variety of physiological activities. Such activities include symbiosis, virulence, motility, antibiotic production, and biofilm formation. Autoinducers come in a number of different forms depending on the species, but the effect that they have is similar in many cases. Autoinducers allow bacteria to communicate both within and between different species. This communication alters gene expression and allows bacteria to mount coordinated responses to their environments, in a manner that is comparable to behavior and signaling in higher organisms. Not surprisingly, it has been suggested that quorum sensing may have been an important evolutionary milestone that ultimately gave rise to multicellular life forms. The term 'autoinduction' was first coined in 1970, when it was observed that the bioluminescent marine bacterium Vibrio fischeri produced a luminescent enzyme (luciferase) only when cultures had reached a threshold population density. At low cell concentrations, V. fischeri did not express the luciferase gene. However, once the cultures had reached exponential growth phase, the luciferase gene was rapidly activated. This phenomenon was termed “autoinduction” because it involved a molecule (autoinducer) that accumulated in a growth medium and induced the synthesis of components of the luminescence system. Subsequent research revealed that the actual autoinducer used by V. fischeri is an acylated homoserine lactone (AHL) signaling molecule. In the most simplified quorum sensing systems, bacteria only need two components to make use of autoinducers. They need a way to produce a signal and a way to respond to that signal. These cellular processes are often tightly coordinated and involve changes in gene expression. The production of autoinducers generally increases as bacterial cell densities increase. Most signals are produced intracellularly and are subsequently secreted in the extracellular environment. Detection of autoinducers often involves diffusion back into cells and binding to specific receptors. Usually, binding of autoinducers to receptors does not occur until a threshold concentration of autoinducers is achieved. Once this has occurred, bound receptors alter gene expression either directly or indirectly. Some receptors are transcription factors themselves, while others relay signals to downstream transcription factors. In many cases, autoinducers participate in forward feedback loops, whereby a small initial concentration of an autoinducer amplifies the production of that same chemical signal to much higher levels. Primarily produced by Gram-negative bacteria, acylated homoserine lactones (AHLs) are a class of small neutral lipid molecules composed of a homoserine lactone ring with an acyl chain. AHLs produced by different species of Gram-negative bacteria vary in the length and composition of the acyl side chain, which often contains 4 to 18 carbon atoms. AHLs are synthesized by AHL synthases. They diffuse in and out of cells by both passive transport and active transport mechanisms. Receptors for AHLs include a number of transcriptional regulators called “R proteins,” which function as DNA binding transcription factors or sensor kinases. Gram-positive bacteria that participate in quorum sensing typically use secreted oligopeptides as autoinducers. Peptide autoinducers usually result from posttranslational modification of a larger precursor molecule. In many Gram-positive bacteria, secretion of peptides requires specialized export mechanisms. For example, some peptide autoinducers are secreted by ATP-binding cassette transporters that couple proteolytic processing and cellular export. Following secretion, peptide autoinducers accumulate in extracellular environments. Once a threshold level of signal is reached, a histidine sensor kinase protein of a two-component regulatory system detects it and a signal is relayed into the cell. As with AHLs, the signal ultimately ends up altering gene expression. Unlike some AHLs, however, most oligopeptides do not act as transcription factors themselves. The free-living bioluminescent marine bacterium, Vibrio harveyi, uses another signaling molecule in addition to an acylated homoserine lactone. This molecule, termed Autoinducer-2 (or AI-2), is a furanosyl borate diester. AI-2, which is also produced and used by a number of Gram-negative and Gram-positive bacteria, is believed to be an evolutionary link between the two major types of quorum sensing circuits. As mentioned, Gram-negative bacteria primarily use acylated homoserine lactones (AHLs) as autoinducer molecules. The minimum quorum sensing circuit in Gram-negative bacteria consists of a protein that synthesizes an AHL and a second, different protein that detects it and causes a change in gene expression. First identified in V. fischeri, these two such proteins are LuxI and LuxR, respectively. Other Gram-negative bacteria use LuxI-like and LuxR-like proteins (homologs), suggesting a high degree of evolutionary conservation. However, among Gram-negatives, the LuxI/LuxI-type circuit has been modified in different species. Described in more detail below, these modifications reflect bacterial adaptations to grow and respond to particular niche environments. Ecologically, V. fischeri is known to have symbiotic associations with a number of eukaryotic hosts, including the Hawaiian Bobtail Squid (Euprymna scolopes). In this relationship, the squid host maintains the bacteria in specialized light organs. The host provides a safe, nutrient rich environment for the bacteria and in turn, the bacteria provide light. Although bioluminescence can be used for mating and other purposes, in E. scolopes it is used for counter illumination to avoid predation. The autoinducer molecule used by V. fischeri is N-(3-oxohexanoyl)-homoserine lactone. This molecule is produced in the cytoplasm by the LuxI synthase enzyme and is secreted through the cell membrane into the extracellular environment. As is true of most autoinducers, the environmental concentration of N-(3-oxohexanoyl)-homoserine lactone is the same as the intracellular concentration within each cell. N-(3-oxohexanoyl)-homoserine lactone eventually diffuses back into cells where it is recognized by LuxR once a threshold concentration (~10 μg/ml) has been reached. LuxR binds the autoinducer and directly activates transcription of the luxICDABE operon. This results in an exponential increase in both the production of autoinducer and in bioluminescence. LuxR bound by autoinducer also inhibits the expression of luxR, which is thought to provide a negative feedback compensatory mechanism to tightly control levels of the bioluminescence genes.