RESUMEN
Allorecognition is a key factor in Dictyostelium development and sociality. It is mediated by two polymorphic transmembrane proteins, TgrB1 and TgrC1, which contain extracellular immunoglobulin domains. TgrB1 and TgrC1 are necessary and sufficient for allorecognition, and they carry out separate albeit overlapping functions in development, but their mechanism of action is unknown. Here, we show that TgrB1 acts as a receptor with TgrC1 as its ligand in cooperative aggregation and differentiation. The proteins bind each other in a sequence-specific manner; TgrB1 exhibits a cell-autonomous function and TgrC1 acts non-cell-autonomously. The TgrB1 cytoplasmic tail is essential for its function and it becomes phosphorylated upon association with TgrC1. Dominant mutations in TgrB1 activate the receptor function and confer partial ligand independence. These roles in development and sociality suggest that allorecognition is crucial in the integration of individual cells into a coherent organism.
Asunto(s)
Adhesión Celular/fisiología , Dictyostelium/metabolismo , Ligandos , Proteínas Protozoarias/metabolismo , Proteínas Portadoras/metabolismo , Diferenciación Celular/fisiología , AMP Cíclico/metabolismo , Proteínas de la Membrana/metabolismo , Mutación/genética , Transporte de Proteínas/fisiología , Proteínas Protozoarias/genéticaRESUMEN
The social amoeba Dictyostelium discoideum integrates into a multicellular organism when individual starving cells aggregate and form a mound. The cells then integrate into defined tissues and develop into a fruiting body that consists of a stalk and spores. Aggregation is initially orchestrated by waves of extracellular cyclic adenosine monophosphate (cAMP), and previous theory suggested that cAMP and other field-wide diffusible signals mediate tissue integration and terminal differentiation as well. Cooperation between cells depends on an allorecognition system comprising the polymorphic adhesion proteins TgrB1 and TgrC1. Binding between compatible TgrB1 and TgrC1 variants ensures that non-matching cells segregate into distinct aggregates prior to terminal development. Here, we have embedded a small number of cells with incompatible allotypes within fields of developing cells with compatible allotypes. We found that compatibility of the allotype encoded by the tgrB1 and tgrC1 genes is required for tissue integration, as manifested in cell polarization, coordinated movement and differentiation into prestalk and prespore cells. Our results show that the molecules that mediate allorecognition in D. discoideum also control the integration of individual cells into a unified developing organism, and this acts as a gating step for multicellularity.
Asunto(s)
Dictyostelium/citología , Regulación del Desarrollo de la Expresión Génica , Complejo GPIb-IX de Glicoproteína Plaquetaria/metabolismo , Proteínas Protozoarias/metabolismo , Alelos , Animales , Adhesión Celular , Comunicación Celular , Diferenciación Celular , Movimiento Celular , Quimiotaxis/fisiología , AMP Cíclico/metabolismo , Dictyostelium/genética , Perfilación de la Expresión Génica , Proteínas Fluorescentes Verdes/metabolismo , Morfogénesis , Unión Proteica , Recombinación Genética , Transcripción GenéticaRESUMEN
Extracellular cAMP functions as a primary ligand for cell surface cAMP receptors throughout Dictyostelium discoideum development, controlling chemotaxis and morphogenesis. The developmental consequences of cAMP signaling and the metabolism of cAMP have been studied in great detail, but it has been unclear how cells export cAMP across the plasma membrane. Here we show pharmacologically and genetically that ABC transporters mediate cAMP export. Using an evolutionary-developmental biology approach, we identified several candidate abc genes and characterized one of them, abcB3, in more detail. Genetic and biochemical evidence suggest that AbcB3 is a component of the cAMP export mechanism in D. discoideum development.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/metabolismo , Quimiotaxis/fisiología , AMP Cíclico/metabolismo , Dictyostelium/crecimiento & desarrollo , Morfogénesis/fisiología , Transportadoras de Casetes de Unión a ATP/genética , Técnicas de Silenciamiento del Gen , Interferencia de ARN , Transducción de Señal/fisiologíaRESUMEN
Data integration procedures combine heterogeneous data sets into predictive models, but they are limited to data explicitly related to the target object type, such as genes. Collage is a new data fusion approach to gene prioritization. It considers data sets of various association levels with the prediction task, utilizes collective matrix factorization to compress the data, and chaining to relate different object types contained in a data compendium. Collage prioritizes genes based on their similarity to several seed genes. We tested Collage by prioritizing bacterial response genes in Dictyostelium as a novel model system for prokaryote-eukaryote interactions. Using 4 seed genes and 14 data sets, only one of which was directly related to the bacterial response, Collage proposed 8 candidate genes that were readily validated as necessary for the response of Dictyostelium to Gram-negative bacteria. These findings establish Collage as a method for inferring biological knowledge from the integration of heterogeneous and coarsely related data sets.
Asunto(s)
Compresión de Datos/métodos , Bases de Datos Genéticas , Dictyostelium/metabolismo , Dictyostelium/microbiología , Bacterias Gramnegativas/fisiología , Proteínas Protozoarias/metabolismo , Proliferación Celular/fisiología , Minería de Datos/métodos , Proteínas Protozoarias/genéticaRESUMEN
Detection of substances tasting bitter to humans occurs in diverse organisms including the social amoeba Dictyostelium discoideum. To establish a molecular mechanism for bitter tastant detection in Dictyostelium, we screened a mutant library for resistance to a commonly used bitter standard, phenylthiourea. This approach identified a G-protein-coupled receptor mutant, grlJ(-), which showed a significantly increased tolerance to phenylthiourea in growth, survival and movement. This mutant was not resistant to a structurally dissimilar potent bitter tastant, denatonium benzoate, suggesting it is not a target for at least one other bitter tastant. Analysis of the cell-signalling pathway involved in the detection of phenylthiourea showed dependence upon heterotrimeric G protein and phosphatidylinositol 3-kinase activity, suggesting that this signalling pathway is responsible for the cellular effects of phenylthiourea. This is further supported by a phenylthiourea-dependent block in the transient cAMP-induced production of phosphatidylinositol (3,4,5)-trisphosphate (PIP3) in wild-type but not grlJ(-) cells. Finally, we have identified an uncharacterized human protein γ-aminobutyric acid (GABA) type B receptor subunit 1 isoform with weak homology to GrlJ that restored grlJ(-) sensitivity to phenylthiourea in cell movement and PIP3 regulation. Our results thus identify a novel pathway for the detection of the standard bitter tastant phenylthiourea in Dictyostelium and implicate a poorly characterized human protein in phenylthiourea-dependent cell responses.
Asunto(s)
Dictyostelium/fisiología , Feniltiourea/química , Fosfatidilinositol 3-Quinasa/genética , Receptores Acoplados a Proteínas G/genética , Receptores de GABA-B/genética , Gusto/fisiología , Movimiento Celular , Supervivencia Celular , AMP Cíclico/metabolismo , Eliminación de Gen , Regulación de la Expresión Génica , Prueba de Complementación Genética , Humanos , Fosfatidilinositol 3-Quinasa/metabolismo , Fosfatos de Fosfatidilinositol/metabolismo , Compuestos de Amonio Cuaternario/química , Receptores Acoplados a Proteínas G/metabolismo , Receptores de GABA-B/metabolismo , Transducción de Señal , Papilas Gustativas/metabolismoRESUMEN
Cooperative social systems are susceptible to cheating by individuals that reap the benefits of cooperation without incurring the costs. There are various theoretical mechanisms for the repression of cheating and many have been tested experimentally. One possibility that has not been tested rigorously is the evolution of mutations that confer resistance to cheating. Here we show that the presence of a cheater in a population of randomly mutated social amoebae can select for cheater-resistance. Furthermore, we show that this cheater-resistance can be a noble strategy because the resister strain does not necessarily exploit other strains. Thus, the evolution of resisters may be instrumental in preserving cooperative behaviour in the face of cheating.
Asunto(s)
Dictyostelium/fisiología , Modelos Biológicos , Conducta Social , Animales , Conducta Cooperativa , Dictyostelium/genética , Evolución Molecular , Genes Protozoarios/genética , Mutación/genética , Proteínas Protozoarias/genética , Proteínas Protozoarias/metabolismo , Esporas Protozoarias/fisiologíaRESUMEN
Cooperation is central to many major transitions in evolution, including the emergence of eukaryotic cells, multicellularity and eusociality. Cooperation can be destroyed by the spread of cheater mutants that do not cooperate but gain the benefits of cooperation from others. However, cooperation can be preserved if cheaters are facultative, cheating others but cooperating among themselves. Several cheater mutants have been studied before, but no study has attempted a genome-scale investigation of the genetic opportunities for cheating. Here we describe such a screen in a social amoeba and show that cheating is multifaceted by revealing cheater mutations in well over 100 genes of diverse types. Many of these mutants cheat facultatively, producing more than their fair share of spores in chimaeras, but cooperating normally when clonal. These findings indicate that phenotypically stable cooperative systems may nevertheless harbour genetic conflicts. The opportunities for evolutionary moves and countermoves in such conflicts may select for the involvement of multiple pathways and numerous genes.
Asunto(s)
Conducta Cooperativa , Dictyostelium/genética , Dictyostelium/fisiología , Mutación/genética , Conducta Social , Amoeba/genética , Amoeba/fisiología , Animales , Agregación Celular , Quimera/genética , Quimera/fisiología , Dictyostelium/citología , Genes Protozoarios/genética , Genoma/genética , Genómica , Myxococcus xanthus/genética , Myxococcus xanthus/fisiología , Fenotipo , Esporas Protozoarias/genética , Esporas Protozoarias/fisiologíaRESUMEN
Classical epistasis analysis can determine the order of function of genes in pathways using morphological, biochemical and other phenotypes. It requires knowledge of the pathway's phenotypic output and a variety of experimental expertise and so is unsuitable for genome-scale analysis. Here we used microarray profiles of mutants as phenotypes for epistasis analysis. Considering genes that regulate activity of protein kinase A in Dictyostelium, we identified known and unknown epistatic relationships and reconstructed a genetic network with microarray phenotypes alone. This work shows that microarray data can provide a uniform, quantitative tool for large-scale genetic network analysis.
Asunto(s)
Dictyostelium/genética , Epistasis Genética , Transcripción Genética , Animales , Proteínas Quinasas Dependientes de AMP Cíclico/genética , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Dictyostelium/enzimología , Mutación , Proteína Quinasa C/genética , Proteína Quinasa C/metabolismoRESUMEN
BACKGROUND: Competitive social interactions are ubiquitous in nature, but their genetic basis is difficult to determine. Much can be learned from single gene knockouts in a eukaryote microbe. The mutants can be competed with the parent to discern the social impact of that specific gene. Dictyostelium discoideum is a social amoeba that exhibits cooperative behavior in the construction of a multicellular fruiting body. It is a good model organism to study the genetic basis of cooperation since it has a sequenced genome and it is amenable to genetic manipulation. When two strains of D. discoideum are mixed, a cheater strain can exploit its social partner by differentiating more spore than its fair share relative to stalk cells. Cheater strains can be generated in the lab or found in the wild and genetic analyses have shown that cheating behavior can be achieved through many pathways. RESULTS: We have characterized the knockout mutant chtB, which was isolated from a screen for cheater mutants that were also able to form normal fruiting bodies on their own. When mixed in equal proportions with parental strain cells, chtB mutants contributed almost 60% of the total number of spores. To do so, chtB cells inhibit wild type cells from becoming spores, as indicated by counts and by the wild type cells' reduced expression of the prespore gene, cotB. We found no obvious fitness costs (morphology, doubling time in liquid medium, spore production, and germination efficiency) associated with the cheating ability of the chtB knockout. CONCLUSIONS: In this study we describe a new gene in D. discoideum, chtB, which when knocked out inhibits the parental strain from producing spores. Moreover, under lab conditions, we did not detect any fitness costs associated with this behavior.
Asunto(s)
Dictyostelium/genética , Dictyostelium/fisiología , Genes Protozoarios , Dictyostelium/citología , Técnicas de Inactivación de Genes , Aptitud Genética , Esporas Protozoarias/citología , Esporas Protozoarias/genéticaRESUMEN
The acyl coenzyme A (CoA) binding protein AcbA is secreted unconventionally and processed into spore differentiation factor 2 (SDF-2), a peptide that coordinates sporulation in Dictyostelium discoideum. We report that AcbA is localized in vesicles that accumulate in the cortex of prespore cells just prior to sporulation. These vesicles are not observed after cells are stimulated to release AcbA but remain visible after stimulation in cells lacking the Golgi reassembly stacking protein (GRASP). Acyl-CoA binding is required for the inclusion of AcbA in these vesicles, and the secretion of AcbA requires N-ethylmaleimide-sensitive factor (NSF). About 1% of the total cellular AcbA can be purified within membrane-bound vesicles. The yield of vesicles decreases dramatically when purified from wild-type cells that were stimulated to release AcbA, whereas the yield from GRASP mutant cells was only modestly altered by stimulation. We suggest that these AcbA-containing vesicles are secretion intermediates and that GRASP functions at a late step leading to the docking/fusion of these vesicles at the cell surface.
Asunto(s)
Dictyostelium/metabolismo , Proteínas Protozoarias/metabolismo , Vesículas Secretoras/metabolismo , Acilcoenzima A/metabolismo , Centrifugación , Detergentes/farmacología , Dictyostelium/citología , Dictyostelium/efectos de los fármacos , Péptidos y Proteínas de Señalización Intercelular , Proteínas Sensibles a N-Etilmaleimida/metabolismo , Péptidos/metabolismo , Unión Proteica/efectos de los fármacos , Transporte de Proteínas/efectos de los fármacos , Vesículas Secretoras/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Ácido gamma-Aminobutírico/farmacologíaRESUMEN
BACKGROUND: Identifying candidate genes in genetic networks is important for understanding regulation and biological function. Large gene expression datasets contain relevant information about genetic networks, but mining the data is not a trivial task. Algorithms that infer Bayesian networks from expression data are powerful tools for learning complex genetic networks, since they can incorporate prior knowledge and uncover higher-order dependencies among genes. However, these algorithms are computationally demanding, so novel techniques that allow targeted exploration for discovering new members of known pathways are essential. RESULTS: Here we describe a Bayesian network approach that addresses a specific network within a large dataset to discover new components. Our algorithm draws individual genes from a large gene-expression repository, and ranks them as potential members of a known pathway. We apply this method to discover new components of the cAMP-dependent protein kinase (PKA) pathway, a central regulator of Dictyostelium discoideum development. The PKA network is well studied in D. discoideum but the transcriptional networks that regulate PKA activity and the transcriptional outcomes of PKA function are largely unknown. Most of the genes highly ranked by our method encode either known components of the PKA pathway or are good candidates. We tested 5 uncharacterized highly ranked genes by creating mutant strains and identified a candidate cAMP-response element-binding protein, yet undiscovered in D. discoideum, and a histidine kinase, a candidate upstream regulator of PKA activity. CONCLUSIONS: The single-gene expansion method is useful in identifying new components of known pathways. The method takes advantage of the Bayesian framework to incorporate prior biological knowledge and discovers higher-order dependencies among genes while greatly reducing the computational resources required to process high-throughput datasets.
Asunto(s)
Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Dictyostelium/enzimología , Expresión Génica , Genómica/métodos , Teorema de Bayes , Dictyostelium/genética , Dictyostelium/crecimiento & desarrollo , Perfilación de la Expresión Génica/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Transducción de Señal/genéticaRESUMEN
Major phenotypic innovations in social amoeba evolution occurred at the transition between the Polysphondylia and group 4 Dictyostelia, which comprise the model organism Dictyostelium discoideum, such as the formation of a new structure, the basal disk. Basal disk differentiation and robust stalk formation require the morphogen DIF-1, synthesized by the polyketide synthase StlB, the des-methyl-DIF-1 methyltransferase DmtA, and the chlorinase ChlA, which are conserved throughout Dictyostelia. To understand how the basal disk and other innovations evolved in group 4, we sequenced and annotated the Polysphondylium violaceum (Pvio) genome, performed cell type-specific transcriptomics to identify cell-type marker genes, and developed transformation and gene knock-out procedures for Pvio. We used the novel methods to delete the Pvio stlB gene. The Pvio stlB- mutants formed misshapen curly sorogens with thick and irregular stalks. As fruiting body formation continued, the upper stalks became more regular, but structures contained 40% less spores. The stlB- sorogens overexpressed a stalk gene and underexpressed a (pre)spore gene. Normal fruiting body formation and sporulation were restored in Pvio stlB- by including DIF-1 in the supporting agar. These data indicate that, although conserved, stlB and its product(s) acquired both a novel role in the group 4 Dictyostelia and a role opposite to that in its sister group.
Asunto(s)
Genoma de Protozoos , Mixomicetos/genética , Mixomicetos/metabolismo , Sintasas Poliquetidas/metabolismo , Proteínas Protozoarias/metabolismo , Mixomicetos/crecimiento & desarrollo , Sintasas Poliquetidas/deficiencia , Sintasas Poliquetidas/genética , Proteínas Protozoarias/genéticaRESUMEN
BACKGROUND: Bioinformatics often leverages on recent advancements in computer science to support biologists in their scientific discovery process. Such efforts include the development of easy-to-use web interfaces to biomedical databases. Recent advancements in interactive web technologies require us to rethink the standard submit-and-wait paradigm, and craft bioinformatics web applications that share analytical and interactive power with their desktop relatives, while retaining simplicity and availability. RESULTS: We have developed dictyExpress, a web application that features a graphical, highly interactive explorative interface to our database that consists of more than 1000 Dictyostelium discoideum gene expression experiments. In dictyExpress, the user can select experiments and genes, perform gene clustering, view gene expression profiles across time, view gene co-expression networks, perform analyses of Gene Ontology term enrichment, and simultaneously display expression profiles for a selected gene in various experiments. Most importantly, these tasks are achieved through web applications whose components are seamlessly interlinked and immediately respond to events triggered by the user, thus providing a powerful explorative data analysis environment. CONCLUSION: dictyExpress is a precursor for a new generation of web-based bioinformatics applications with simple but powerful interactive interfaces that resemble that of the modern desktop. While dictyExpress serves mainly the Dictyostelium research community, it is relatively easy to adapt it to other datasets. We propose that the design ideas behind dictyExpress will influence the development of similar applications for other model organisms.
Asunto(s)
Biología Computacional/métodos , Bases de Datos Genéticas , Dictyostelium/genética , Perfilación de la Expresión Génica/métodos , Internet , Programas Informáticos , Animales , Interfaz Usuario-ComputadorRESUMEN
William Farnsworth Loomis studied the social amoeba Dictyostelium discoideum for more than fifty years as a professor of biology at the University of California, San Diego, USA. This biographical reflection describes Dr. Loomis' major scientific contributions to the field within a career arc that spanned the early days of molecular biology up to the present day where the acquisition of high-dimensional datasets drive research. Dr. Loomis explored the genetic control of social amoeba development, delineated mechanisms of cell differentiation, and significantly advanced genetic and genomic technology for the field. The details of Dr. Loomis' multifaceted career are drawn from his published work, from an autobiographical essay that he wrote near the end of his career and from extensive conversations between him and the two authors, many of which took place on the deck of his beachfront home in Del Mar, California.
Asunto(s)
Dictyostelium/genética , Dictyostelium/fisiología , Diferenciación Celular , Biología Evolutiva/historia , Genómica , Historia del Siglo XX , Historia del Siglo XXI , Modelos Genéticos , MutagénesisRESUMEN
Both animals and amoebae use phagocytosis and DNA-based extracellular traps as anti-bacterial defense mechanisms. Whether, like animals, amoebae also use tissue-level barriers to reduce direct contact with bacteria has remained unclear. We have explored this question in the social amoeba Dictyostelium discoideum, which forms plaques on lawns of bacteria that expand as amoebae divide and bacteria are consumed. We show that CadA, a cell adhesion protein that functions in D. discoideum development, is also a bacterial agglutinin that forms a protective interface at the plaque edge that limits exposure of vegetative amoebae to bacteria. This interface is important for amoebal survival when bacteria-to-amoebae ratios are high, optimizing amoebal feeding behavior, and protecting amoebae from oxidative stress. Lectins also control bacterial access to the gut epithelium of mammals to limit inflammatory processes; thus, this strategy of antibacterial defense is shared across a broad spectrum of eukaryotic taxa.
Asunto(s)
Moléculas de Adhesión Celular/genética , Dictyostelium/genética , Inflamación/genética , Lectinas/genética , Aglutinación/genética , Aglutininas/genética , Animales , Bacillus subtilis/genética , Bacillus subtilis/patogenicidad , Dictyostelium/microbiología , Interacciones Huésped-Patógeno/genética , Inflamación/microbiología , Mamíferos/microbiología , Mamíferos/parasitología , Micrococcus luteus/genética , Micrococcus luteus/patogenicidad , Fagocitosis/genética , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidadRESUMEN
Social amoebae and humans use common strategies to orchestrate their interactions with the bacteria in their respective environments and within their bodies. These strategies include the elimination of bacteria by phagocytosis, the establishment of mutualistic interactions, the elaboration of physical barriers, and the deployment of innate immune cells. Many of the molecular mechanisms that humans and social amoebae employ differ, but there are striking similarities that may inform studies in each organism. In this topical review we highlight the similarities and consider what we might learn by comparing these highly divergent species. We focus on recent work in Dictyostelium discoideum with hopes of stimulating work in this area and with the expectation that new mechanistic details uncovered in social amoebae-bacteria interactions will inform microbiome management in humans.
Asunto(s)
Dictyostelium/microbiología , Dictyostelium/fisiología , Microbiota , Animales , Bacterias , Trampas Extracelulares/metabolismo , Humanos , Intestinos/microbiología , Lectinas/metabolismo , Ratones , Fagocitosis , Suelo , SimbiosisRESUMEN
The eukaryotic amoeba Dictyostelium discoideum is commonly used to study sociality. The amoebae cooperate during development, exhibiting altruism, cheating, and kin-discrimination, but growth while preying on bacteria has been considered asocial. Here we show that Dictyostelium are cooperative predators. Using mutants that grow poorly on Gram-negative bacteria but grow well on Gram-positive bacteria, we show that growth depends on cell-density and on prey type. We also found synergy, by showing that pairwise mixes of different mutants grow well on live Gram-negative bacteria. Moreover, wild-type amoebae produce diffusible factors that facilitate mutant growth and some mutants exploit the wild type in mixed cultures. Finding cooperative predation in D. discoideum should facilitate studies of this fascinating phenomenon, which has not been amenable to genetic analysis before.
Asunto(s)
Dictyostelium/fisiología , Animales , Dictyostelium/genética , Dictyostelium/crecimiento & desarrollo , Genes Protozoarios , Klebsiella pneumoniae , Mutación , Fagocitosis , Conducta Predatoria , Esporas Protozoarias , Staphylococcus aureusRESUMEN
The social amoeba Dictyostelium discoideum maintains a microbiome during multicellular development; bacteria are carried in migrating slugs and as endosymbionts within amoebae and spores. Bacterial carriage and endosymbiosis are induced by the secreted lectin discoidin I that binds bacteria, protects them from extracellular killing, and alters their retention within amoebae. This altered handling of bacteria also occurs with bacteria coated by plant lectins and leads to DNA transfer from bacteria to amoebae. Thus, lectins alter the cellular response of D. discoideum to bacteria to establish the amoebae's microbiome. Mammalian cells can also maintain intracellular bacteria when presented with bacteria coated with lectins, so heterologous lectins may induce endosymbiosis in animals. Our results suggest that endogenous or environmental lectins may influence microbiome homeostasis across eukaryotic phylogeny.
Asunto(s)
Dictyostelium/metabolismo , Dictyostelium/microbiología , Discoidinas/metabolismo , Klebsiella pneumoniae/fisiología , Microbiota/fisiología , Transporte Biológico , ADN Bacteriano/metabolismo , Klebsiella pneumoniae/metabolismo , SimbiosisRESUMEN
The Dictyostelium protein kinase YakA is required for the growth-to-development transition. During growth YakA controls the cell cycle, regulating the intervals between cell divisions. When starved for nutrients Dictyostelium cells arrest growth and undergo changes in gene expression, decreasing vegetative mRNAs and inducing the expression of pkaC. YakA is an effector of these changes, being necessary for the decrease of vegetative mRNA expression and the increase of protein kinase A (PKA) activity that will ultimately regulate expression of adenylyl cyclase, cAMP synthesis, and the induction of development. We report a role for this kinase in the response to nitrosoative or oxidative stress of Dictyostelium cells. Hydrogen peroxide and sodium nitroprusside arrest the growth of cells and trigger cAMP synthesis and activation of PKA in a manner similar to the well-established response to nutrient starvation. We have found that yakA null cells are hypersensitive to nitrosoative/oxidative stress and that a second-site mutation in pkaC suppresses this sensitivity. The response to different stresses has been investigated and YakA, cAMP, and PKA have been identified as components of the pathway that regulate the growth arrest that follows treatment with compounds that generate reactive oxygen species. The effect of different types of stress was evaluated in Dictyostelium and the YakA/PKA pathway was also implicated in the response to heat stress.
Asunto(s)
Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Dictyostelium/metabolismo , Estrés Oxidativo , Proteínas Quinasas/metabolismo , Espermina/análogos & derivados , Animales , División Celular/fisiología , Dictyostelium/efectos de los fármacos , Regulación de la Expresión Génica , Calor/efectos adversos , Peróxido de Hidrógeno/farmacología , Óxido Nítrico/metabolismo , Donantes de Óxido Nítrico/farmacología , Óxidos de Nitrógeno , Nitroprusiato/farmacología , Oxidantes/farmacología , Subunidades de Proteína/metabolismo , Espermina/farmacologíaRESUMEN
The Amoebozoa are a sister clade to the fungi and the animals, but are poorly sampled for completely sequenced genomes. The social amoeba Dictyostelium discoideum and amitochondriate pathogen Entamoeba histolytica are the first Amoebozoa with genomes completely sequenced. Both organisms are classified under the Conosa subphylum. To identify Amoebozoa-specific genomic elements, we compared these two genomes to each other and to other eukaryotic genomes. An expanded phylogenetic tree built from the complete predicted proteomes of 23 eukaryotes places the two amoebae in the same lineage, although the divergence is estimated to be greater than that between animals and fungi, and probably happened shortly after the Amoebozoa split from the opisthokont lineage. Most of the 1,500 orthologous gene families shared between the two amoebae are also shared with plant, animal, and fungal genomes. We found that only 42 gene families are distinct to the amoeba lineage; among these are a large number of proteins that contain repeats of the FNIP domain, and a putative transcription factor essential for proper cell type differentiation in D. discoideum. These Amoebozoa-specific genes may be useful in the design of novel diagnostics and therapies for amoebal pathologies.