Sporulation in Bacillus subtilis

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Fig1. The sporulation process of Bacillus subtilis Sporulation in Bacillus subtilis.jpg
Fig1. The sporulation process of Bacillus subtilis

Bacillus subtilis is a rod-shaped, Gram-positive bacteria that is naturally found in soil and vegetation, and is known for its ability to form a small, tough, protective and metabolically dormant endospore. B. subtilis can divide symmetrically to make two daughter cells (binary fission), or asymmetrically, producing a single endospore that is resistant to environmental factors such as heat, desiccation, radiation and chemical insult which can persist in the environment for long periods of time. The endospore is formed at times of nutritional stress, allowing the organism to persist in the environment until conditions become favourable. The process of endospore formation has profound morphological and physiological consequences: radical post-replicative remodelling of two progeny cells, accompanied eventually by cessation of metabolic activity in one daughter cell (the spore) and death by lysis of the other (the ‘mother cell’).

Contents

Overview

Commitment to sporulation

Although sporulation in B. subtilis is induced by starvation, the sporulation developmental program is not initiated immediately when growth slows due to nutrient limitation. A variety of alternative responses can occur, including the activation of flagellar motility to seek new food sources by chemotaxis, the production of antibiotics to destroy competing soil microbes, the secretion of hydrolytic enzymes to scavenge extracellular proteins and polysaccharides, or the induction of ‘competence’ for uptake of exogenous DNA for consumption, with the occasional side-effect that new genetic information is stably integrated. Sporulation is the last-ditch response to starvation and is suppressed until alternative responses prove inadequate. Even then, certain conditions must be met such as chromosome integrity, the state of chromosomal replication, and the functioning of the Krebs cycle. [1]

Nature of regulation

Sporulation requires a great deal of time and also a lot of energy and is essentially irreversible, [2] making it crucial for a cell to monitor its surroundings efficiently and ensure that sporulation is embarked upon at only the most appropriate times. The wrong decision can be catastrophic: a vegetative cell will die if the conditions are too harsh, while bacteria forming spores in an environment which is conducive to vegetative growth will be out competed. [3] In short, initiation of sporulation is a very tightly regulated network with numerous checkpoints for efficient control.[ citation needed ]

Control points

Two transcriptional regulators, σH and Spo0A, play key roles in initiation of sporulation. Several additional proteins participate, mainly by controlling the accumulated concentration of Spo0A~P. Spo0A lies at the end of a series of inter-protein phosphotransfer reactions, Kin–Spo0F–Spo0B–Spo0A, termed as a ‘phosphorelay’. The regulation of these various factors controlling the accumulated concentration of Spo0A~P, and their interactions are described in detail in Figure2:

Fig2. Gene Regulatory Network for Initiation of Sporulation in Bacillus subtilis Gene Regulatory Network for Initiation of Sporulation in Bacillus subtilis.png
Fig2. Gene Regulatory Network for Initiation of Sporulation in Bacillus subtilis

In the table below, the term 'Activators' refers to genes/proteins that ultimately result in initiation of sporulation and 'Repressors' refers to the ones that inhibit this initiation of sporulation.

ActivatorsRegulationRepressorsRegulation
KinATransfers phosphate to the Spo0FSdaBlocks autophosphorylation of KinA
KinBTransfers phosphate to the Spo0FKipIBlocks autophosphorylation of KinA
Spo0AActivates several key sporulation-specific genesSpo0ANegatively controls transcription of abrB
Spo0H (σH)Activates phrE geneComAActivates competence
Spo0BPhosphotransferase initiationSinRNegative regulation of kinB
Spo0FPhosphotransferase initiationRapA, RapB, RapE, and RapHDephosphorylation of Spo0F~P
ComAActivates phrASpo0E, YisI, and YnzDDephosphorylation of Spo0A~P
SinIAntagonist of SinRGTP-bound CodYInhibits rapA-phrA
KipAInhibits kipI geneAbrBEncodes a repressor of spo0H
PhrASuppresses dephosphorylation activity of RapAHprOverexpression inhibits sporulation
PhrEInhibits RapEDnaAOver expression of Sda
PhrHInhibits RapH

Homologous networks in eukaryotes

Spores form a part of the life cycles of a diverse range of organisms such as many bacteria, plants, algae, fungi and some protozoa. In Saccharomyces cerevisiae (Kingdom Fungi), the set of early genes activating sporulation is induced by Ime1 (Inducer of Meiosis 1) and a regulator of middle genes is Ndt80p. [4] Similarly, in Dictyostelium discoideum (slime mold), SDF-2 and cytokinins are secreted during late development to trigger signal transduction pathways that lead to an increase in the activity of the cAMP-dependent protein kinase, PKA, which triggers rapid encapsulation as well as ensuring spore dormancy. [5]

See also

Related Research Articles

<i>Bacillus</i> Genus of bacteria

Bacillus is a genus of Gram-positive, rod-shaped bacteria, a member of the phylum Bacillota, with 266 named species. The term is also used to describe the shape (rod) of other so-shaped bacteria; and the plural Bacilli is the name of the class of bacteria to which this genus belongs. Bacillus species can be either obligate aerobes which are dependent on oxygen, or facultative anaerobes which can survive in the absence of oxygen. Cultured Bacillus species test positive for the enzyme catalase if oxygen has been used or is present.

Autolysins are endogenous lytic enzymes that break down the peptidoglycan components of biological cells which enables the separation of daughter cells following cell division. They are involved in cell growth, cell wall metabolism, cell division and separation, as well as peptidoglycan turnover and have similar functions to lysozymes.

<span class="mw-page-title-main">Endospore</span> Protective structure formed by bacteria

An endospore is a dormant, tough, and non-reproductive structure produced by some bacteria in the phylum Bacillota. The name "endospore" is suggestive of a spore or seed-like form, but it is not a true spore. It is a stripped-down, dormant form to which the bacterium can reduce itself. Endospore formation is usually triggered by a lack of nutrients, and usually occurs in gram-positive bacteria. In endospore formation, the bacterium divides within its cell wall, and one side then engulfs the other. Endospores enable bacteria to lie dormant for extended periods, even centuries. There are many reports of spores remaining viable over 10,000 years, and revival of spores millions of years old has been claimed. There is one report of viable spores of Bacillus marismortui in salt crystals approximately 250 million years old. When the environment becomes more favorable, the endospore can reactivate itself into a vegetative state. Most types of bacteria cannot change to the endospore form. Examples of bacterial species that can form endospores include Bacillus cereus, Bacillus anthracis, Bacillus thuringiensis, Clostridium botulinum, and Clostridium tetani.

<span class="mw-page-title-main">Transformation (genetics)</span> Genetic alteration of a cell by uptake of genetic material from the environment

In molecular biology and genetics, transformation is the genetic alteration of a cell resulting from the direct uptake and incorporation of exogenous genetic material from its surroundings through the cell membrane(s). For transformation to take place, the recipient bacterium must be in a state of competence, which might occur in nature as a time-limited response to environmental conditions such as starvation and cell density, and may also be induced in a laboratory.

<i>Bacillus subtilis</i> Catalase-positive bacterium

Bacillus subtilis, known also as the hay bacillus or grass bacillus, is a Gram-positive, catalase-positive bacterium, found in soil and the gastrointestinal tract of ruminants, humans and marine sponges. As a member of the genus Bacillus, B. subtilis is rod-shaped, and can form a tough, protective endospore, allowing it to tolerate extreme environmental conditions. B. subtilis has historically been classified as an obligate aerobe, though evidence exists that it is a facultative anaerobe. B. subtilis is considered the best studied Gram-positive bacterium and a model organism to study bacterial chromosome replication and cell differentiation. It is one of the bacterial champions in secreted enzyme production and used on an industrial scale by biotechnology companies.

"Candidatus Epulonipiscium" is a genus of Gram-positive bacteria that have a symbiotic relationship with surgeonfish. These bacteria are known for their unusually large size, many ranging from .2 - .7 mm in length. Until the discovery of Thiomargarita namibiensis in 1999, Epulonipiscium species were thought to be the largest bacteria. They are still the largest known heterotrophic bacteria.

<span class="mw-page-title-main">Lysogenic cycle</span> Process of virus reproduction

Lysogeny, or the lysogenic cycle, is one of two cycles of viral reproduction. Lysogeny is characterized by integration of the bacteriophage nucleic acid into the host bacterium's genome or formation of a circular replicon in the bacterial cytoplasm. In this condition the bacterium continues to live and reproduce normally, while the bacteriophage lies in a dormant state in the host cell. The genetic material of the bacteriophage, called a prophage, can be transmitted to daughter cells at each subsequent cell division, and later events can release it, causing proliferation of new phages via the lytic cycle. Lysogenic cycles can also occur in eukaryotes, although the method of DNA incorporation is not fully understood. For instance the AIDS viruses can either infect humans lytically, or lay dormant (lysogenic) as part of the infected cells' genome, keeping the ability to return to lysis at a later time. The rest of this article is about lysogeny in bacterial hosts.

<i>Myxococcus xanthus</i> Slime bacterium

Myxococcus xanthus is a gram-negative, rod-shaped species of myxobacteria that exhibits various forms of self-organizing behavior in response to environmental cues. Under normal conditions with abundant food, it exists as a predatory, saprophytic single-species biofilm called a swarm. Under starvation conditions, it undergoes a multicellular development cycle.

<span class="mw-page-title-main">Spo11</span>

Spo11 is a protein that in humans is encoded by the SPO11 gene. Spo11, in a complex with mTopVIB, creates double strand breaks to initiate meiotic recombination. Its active site contains a tyrosine which ligates and dissociates with DNA to promote break formation. One Spo11 protein is involved per strand of DNA, thus two Spo11 proteins are involved in each double stranded break event.

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.

<i>Dictyostelium discoideum</i> Species of slime mould

Dictyostelium discoideum is a species of soil-dwelling amoeba belonging to the phylum Amoebozoa, infraphylum Mycetozoa. Commonly referred to as slime mold, D. discoideum is a eukaryote that transitions from a collection of unicellular amoebae into a multicellular slug and then into a fruiting body within its lifetime. Its unique asexual lifecycle consists of four stages: vegetative, aggregation, migration, and culmination. The lifecycle of D. discoideum is relatively short, which allows for timely viewing of all stages. The cells involved in the lifecycle undergo movement, chemical signaling, and development, which are applicable to human cancer research. The simplicity of its lifecycle makes D. discoideum a valuable model organism to study genetic, cellular, and biochemical processes in other organisms.

Spore photoproduct lyase is a radical SAM enzyme that repairs DNA cross linking of thymine bases caused by UV-radiation. There are several types of thymine cross linking, but SPL specifically targets 5-thyminyl-5,6-dihydrothymine, which is also called spore photoproduct (SP). Spore photoproduct is the predominant type of thymine crosslinking in germinating endospores, which is why SPL is unique to organisms that produce endospores, such as Bacillus subtilis. Other types of thymine crosslinking, such as cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimidone photoproducts (6-4PPs), are less commonly formed in endospores. These differences in DNA crosslinking are a function of differing DNA structure. Spore genomic DNA features many DNA binding proteins called small acid soluble proteins, which changes the DNA from the traditional B-form conformation to an A-from conformation. This difference in conformation is believed to be the reason why dormant spores predominantly accumulate SP in response to UV-radiation, rather than other forms of cross linking. Spores cannot repair cross-linking while dormant, instead the SPs are repaired during germination to allow the vegetative cell to function normally. When not repaired, spore photoproduct and other types of crosslinking can cause mutations by blocking transcription and replication past the point of the crosslinking. The repair mechanism utilizing spore photoproduct lyase is one of the reasons for the resilience of certain bacterial spores.

<i>Bacillus anthracis</i> Species of bacterium

Bacillus anthracis is a gram-positive and rod-shaped bacterium that causes anthrax, a deadly disease to livestock and, occasionally, to humans. It is the only permanent (obligate) pathogen within the genus Bacillus. Its infection is a type of zoonosis, as it is transmitted from animals to humans. It was discovered by a German physician Robert Koch in 1876, and became the first bacterium to be experimentally shown as a pathogen. The discovery was also the first scientific evidence for the germ theory of diseases.

<span class="mw-page-title-main">SpoT</span>

Bifunctional (p)ppGpp synthase/hydrolase SpoT or SpoT is a regulatory enzyme in the RelA/SpoT Homologue (RSH) protein family that synthesizes and hydrolyzes (p)ppGpp to regulate the bacterial stringent response to environmental stressors. SpoT is considered a "long" form RSH protein and is found in many bacteria and plant chloroplasts. SpoT and its homologues have been studied in bacterial model organism E.coli for their role in the production and degradation of (p)ppGpp in the stringent response pathway.

<span class="mw-page-title-main">Meiotic recombination checkpoint</span>

The meiotic recombination checkpoint monitors meiotic recombination during meiosis, and blocks the entry into metaphase I if recombination is not efficiently processed.

<span class="mw-page-title-main">Sda protein domain</span>

In molecular biology, the protein domain Sda is short for suppressor of dnaA or otherwise known as sporulation inhibitor A. It is found only in bacteria. This protein domain is highly important to cell survival. When starved of nutrients, the cell is under extreme stress so undergoes a series of reactions to increase the chances of survival. One method is to form endospores which can withstand a large amount of environmental pressure. Sda protein domain is a checkpoint which prevents the formation of spores. The Sda domain affects cell signalling. It prevents the cell communicating the stress that it is under, which is crucial if the cell is to survive.

<span class="mw-page-title-main">Endospore staining</span>

Endospore staining is a technique used in bacteriology to identify the presence of endospores in a bacterial sample. Within bacteria, endospores are protective structures used to survive extreme conditions, including high temperatures making them highly resistant to chemicals. Endospores contain little or no ATP which indicates how dormant they can be. Endospores contain a tough outer coating made up of keratin which protects them from nucleic DNA as well as other adaptations. Endospores are able to regerminate into vegetative cells, which provides a protective nature that makes them difficult to stain using normal techniques such as simple staining and gram staining. Special techniques for endospore staining include the Schaeffer–Fulton stain and the Moeller stain.

SpoIVB peptidase is an enzyme. This enzyme catalyses the following chemical reaction

<span class="mw-page-title-main">Histidine phosphotransfer domain</span>

Histidine phosphotransfer domains and histidine phosphotransferases are protein domains involved in the "phosphorelay" form of two-component regulatory systems. These proteins possess a phosphorylatable histidine residue and are responsible for transferring a phosphoryl group from an aspartate residue on an intermediate "receiver" domain, typically part of a hybrid histidine kinase, to an aspartate on a final response regulator.

The Monovalent Cation:Proton Antiporter-2 (CPA2) Family is a moderately large family of transporters belonging to the CPA superfamily. Members of the CPA2 family have been found in bacteria, archaea and eukaryotes. The proteins of the CPA2 family consist of between 333 and 900 amino acyl residues and exhibit 10-14 transmembrane α-helical spanners (TMSs).

References

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