RESUMO
Malaria is one of the deadliest vector-borne diseases in the world. Researchers are developing new genetic and conventional vector control strategies to attempt to limit its burden. Novel control strategies require detailed safety assessment to ensure responsible and successful deployments. Anopheles gambiae sensu stricto (s.s.) and Anopheles coluzzii, two closely related subspecies within the species complex Anopheles gambiae sensu lato (s.l.), are among the dominant malaria vectors in sub-Saharan Africa. These two subspecies readily hybridise and compete in the wild and are also known to have distinct niches, each with spatially and temporally varying carrying capacities driven by precipitation and land use factors. We model the spread and persistence of a population-modifying gene drive system in these subspecies across sub-Saharan Africa by simulating introductions of genetically modified mosquitoes across the African mainland and its offshore islands. We explore transmission of the gene drive between the two subspecies that arise from different hybridisation mechanisms, the effects of both local dispersal and potential wind-aided migration to the spread, and the development of resistance to the gene drive. Given the best current available knowledge on the subspecies' life histories, we find that an introduced gene drive system with typical characteristics can plausibly spread from even distant offshore islands to the African mainland with the aid of wind-driven migration, with resistance beginning to take over within a decade. Our model accounts for regional to continental scale mechanisms, and demonstrates a range of realistic dynamics including the effect of prevailing wind on spread and spatio-temporally varying carrying capacities for subspecies. As a result, it is well-placed to answer future questions relating to mosquito gene drives as important life history parameters become better understood.
Assuntos
Anopheles , Malária , África Subsaariana , Animais , Anopheles/genética , Controle de Mosquitos , Mosquitos Vetores/genética , Dinâmica PopulacionalRESUMO
The series of responses in this article were gathered as part of an online mini conference held in September 2021 that sought to explore different ideas and articulations of school autonomy reform across the world (Australia, Canada, England, Ireland, the USA, Norway, Sweden and New Zealand). It centred upon an important question: what needs to happen for school autonomy to be mobilised to create more equitable public schools and systems of education? There was consensus across the group that school autonomy reform creates further inequities at school and system levels when driven by the logics of marketisation, competition, economic efficiency and public accountability. Against the backdrop of these themes, the conference generated discussion and debate where provocations and points of agreement and disagreement about issues of social justice and the mobilisation of school autonomy reform were raised. As an important output of this discussion, we asked participants to write a short response to the guiding conference question. The following are these responses which range from philosophical considerations, systems and governance perspectives, national particularities and teacher and principal perspectives.
RESUMO
The geographic niches of many species are dramatically changing as a result of environmental and anthropogenic impacts such as global climate change and the introduction of invasive species. In particular, genetically compatible subspecies that were once geographically separated are being reintroduced to one another. This is of concern for conservation, where rare or threatened subspecies could be bred out by hybridising with their more common relatives, and for commercial interests, where the stock or quality of desirable harvested species could be compromised. It is also relevant to disease ecology, where disease transmission is heterogeneous among subspecies and hybridisation may affect the rate and spatial spread of disease. We develop and investigate a mathematical model to combine competitive effects via the Lotka-Volterra model with hybridisation effects via mate choice. The species complex is structured into two classes: a subspecies of interest (named x), and other subspecies including any hybrids produced (named y). We show that in the absence of limit cycles the model has four possible equilibrium outcomes, representing every combination: total extinction, x-dominance (y extinct), y-dominance (x extinct), and at most a single coexistence equilibrium. We give conditions for which limit cycles cannot exist, then further show that the "total extinction" equilibrium is always unstable, that y-dominance is always stable, and that the other equilibria have stability depending on the model parameters. We demonstrate that both x-dominance and coexistence are achievable under a wide range of parameter values and initial conditions, which corresponds with empirical evidence of known competing-hybridising systems. We then briefly examine bifurcation behaviour. In particular, we note that a subcritical bifurcation is possible in which a "catastrophic" transition from x-dominance to y-dominance can occur, representing an invasion event. Finally, we briefly examine the common complication of time-varying carrying capacity, showing that such a case can make coexistence more likely.
Assuntos
Ecossistema , Modelos Biológicos , Conservação dos Recursos Naturais , Ecologia , Espécies IntroduzidasRESUMO
Phosphoinositides play important roles in regulating the cytoskeleton and vesicle trafficking, potentially important processes at the cleavage furrow. However, it remains unclear which, if any, of the phosphoinositides play a role during cytokinesis. A systematic analysis to determine if any of the phosphoinositides might be present or of functional importance at the cleavage furrow has not been published. Several studies hint at a possible role for one or more phosphoinositides at the cleavage furrow. The best of these are genetic data identifying mutations in phosphoinositide-modifying enzymes (a PtdIns(4)P-5-kinase in S. pombe and a PI-4-kinase in D. melanogaster) that interfere with cytokinesis. The genetic nature of these experiments leaves questions as to how direct may be their contribution to cytokinesis. Here we show that a single phosphoinositide, PtdIns(4,5)P2, specifically accumulates at the furrow. Interference with PtdIns(4,5)P2 interferes with adhesion of the plasma membrane to the contractile ring at the furrow. Finally, four distinct interventions to specifically interfere with PtdIns(4,5)P2 each impair cytokinesis. We conclude that PtdIns(4,5)P2 is present at the cleavage furrow and is required for normal cytokinesis at least in part because of a role in adhesion between the contractile ring and the plasma membrane.
Assuntos
Citocinese/fisiologia , Fosfatos de Fosfatidilinositol/metabolismo , Actinas/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Animais , Células CHO , Membrana Celular/metabolismo , Cricetinae , Cricetulus , Vetores Genéticos , Proteínas de Fluorescência Verde/metabolismo , Células HeLa , Humanos , Camundongos , Células NIH 3T3 , Fosfatidilinositol 4,5-Difosfato , Fosfolipase C gama/metabolismo , Proteínas/metabolismoRESUMO
The recently identified RhoBTB family is a member of the Rho GTPase family. One family member, RhoBTB2, has been implicated as a tumor suppressor in lung and breast cancer. Studies have shown that RhoBTB2 binds to the ubiquitin ligase scaffold Cul3 and that Cul3 regulates RhoBTB2 protein levels by ubiquitinating RhoBTB2 directly, leading to its degradation by the proteasome. This chapter details the cell biological and biochemical methods for analyzing the regulation of RhoBTB2 by Cul3.
Assuntos
Proteínas Culina/fisiologia , Proteínas de Ligação ao GTP/fisiologia , Proteínas Supressoras de Tumor/fisiologia , Animais , Anticorpos/farmacologia , Linhagem Celular , Proteínas de Ligação ao GTP/imunologia , Cabras , Células HeLa , Humanos , Complexos Multiproteicos/isolamento & purificação , Coelhos , Proteínas Supressoras de Tumor/imunologia , UbiquitinaçãoRESUMO
Phosphatidylinositol 4-phosphate 5-kinases [PtdIns4P5Ks] synthesise the majority of cellular phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)] and phospholipase D1 (PLD1) synthesises large amounts of phosphatidic acid (PtdOH). The activities of PtdIns4P5Ks and PLDs are thought to be coupled during cell signalling in order to support large simultaneous increases in both PtdIns(4,5)P(2) and PtdOH, since PtdOH activates PtdIns4P5Ks and PLD1 requires PtdIns(4,5)P(2) as a cofactor. However, little is known about the control of such a system. Membrane recruitment of ADP-ribosylation factors (Arfs) activates both PtdIns4P5Ks and PLDs, but it is not known if each enzyme is controlled in series by different Arfs or in parallel by a single form. We show through pull-down and vesicle sedimentation interaction assays that PtdIns4P5K activation may be facilitated by Arf-enhanced membrane association. However PtdIns4P5Ks discriminate poorly between near homogeneously myristoylated Arf1 and Arf6 although examples of all three known active isoforms (mouse alpha>beta, gamma) respond to these G-proteins. Conversely PLD1 genuinely prefers Arf1 and so the two lipid metabolising enzymes are differentially controlled. We propose that isoform selective Arf/PLD interaction and not Arf/PtdIns4P5K will be the critical trigger in the formation of distinct, optimal triples of Arf/PLDs/PtdIns4P5Ks and be the principle regulator of any coupled increases in the signalling lipids PtdIns(4,5)P(2) and PtdOH.
Assuntos
Fator 1 de Ribosilação do ADP/metabolismo , Fatores de Ribosilação do ADP/metabolismo , Fosfolipase D/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Fator 1 de Ribosilação do ADP/química , Fator 1 de Ribosilação do ADP/genética , Fator 6 de Ribosilação do ADP , Fatores de Ribosilação do ADP/química , Fatores de Ribosilação do ADP/genética , Animais , Sequência de Bases , Sistema Livre de Células , Primers do DNA/genética , Ativação Enzimática , Guanosina Trifosfato/metabolismo , Células HL-60 , Humanos , Técnicas In Vitro , Cinética , Lipídeos de Membrana/metabolismo , Camundongos , Modelos Biológicos , Ácidos Mirísticos/química , Fosfolipase D/genética , Fosfolipídeos/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transdução de SinaisRESUMO
BACKGROUND: Crohn's disease is an autoimmune inflammatory disorder of the gastrointestinal tract and is characterized clinically by dysregulation of both pro-inflammatory and anti-inflammatory cytokine signaling networks. The function of the Crohn's disease protein, NOD2, highlights the biphasic nature of the pathology of Crohn's disease. NOD2 can both strongly activate and negatively attenuate NF-kB signaling. The biochemical mechanism for this dual function of NOD2 is unknown. RESULTS: We demonstrate that NOD2 activation leads to ubiquitinylation of NEMO, a key component of the NF-kB signaling complex. This ubiquitinylation is agonist dependant, and it does not regulate proteosomal destruction of NEMO. We show the NOD2-dependent ubiquitinylation of NEMO is dependent on the scaffolding protein kinase RIP2. Crohn's disease-associated polymorphisms of NOD2 show a decreased ability to bind RIP2, and this decreased ability to bind RIP2 correlates with a decreased ability to ubiquitinylate NEMO. We map the site of NEMO ubiquitinylation to a novel NEMO ubiquitinylation site (Lysine 285) and show that this ubiquityinylation occurs in vivo. Lastly, we show functionally that RIP2-induced ubiquitinylation of NEMO is at least in part responsible for RIP2-mediated NF-kB activation. CONCLUSIONS: These data suggest that this novel mode of regulation of the NF-kB signaling pathway could be a factor underlying the pathogenesis of Crohn's disease.
Assuntos
Proteínas de Transporte/metabolismo , Doença de Crohn/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Transdução de Sinais/fisiologia , Ubiquitina/metabolismo , Sequência de Aminoácidos , Western Blotting , Células Cultivadas , Humanos , Quinase I-kappa B , Imunoprecipitação , Peptídeos e Proteínas de Sinalização Intracelular/genética , Espectrometria de Massas , Dados de Sequência Molecular , NF-kappa B/metabolismo , Proteína Adaptadora de Sinalização NOD2 , Polimorfismo Genético , Proteína Serina-Treonina Quinase 2 de Interação com Receptor , Alinhamento de SequênciaRESUMO
A community-based study of human immunodeficiency virus type 2 (HIV-2) infection was conducted in a rural village in northern Guinea Bissau, West Africa to assess the relationship between plasma HIV-2 RNA levels, CD4 lymphocyte percentage, and survival over an 8-year period. The cohort of 133 HIV-2-infected individuals and 160 HIV-uninfected controls enrolled in 1991 were followed up at home until 1998. Thirty-one (23%) HIV-2-infected and 24 (16%) HIV-uninfected individuals died over the follow-up period (mortality hazard ratio 1.7, 95% CI 1.0, 2.9; p= 0.06). In HIV-2-infected individuals, the median HIV-2 RNA level was 347 copies/ml and the mean CD4% was 28.6. Both plasma viremia and CD4% were independent predictors of survival, with hazard ratios increasing by 1.6 (95% CI, 1.1, 2.3) for each log(10) increase of plasma viremia and 1.7 (1.1, 2.6) for each 10% decrease of CD4%. Infected subjects with a plasma viral load >or= the median (347 copies/ml) and a CD4% Assuntos
Contagem de Linfócito CD4
, Infecções por HIV/fisiopatologia
, HIV-2/isolamento & purificação
, Taxa de Sobrevida
, Carga Viral
, Adulto
, Idoso
, Estudos de Casos e Controles
, Feminino
, Guiné-Bissau
, Infecções por HIV/imunologia
, Infecções por HIV/virologia
, Humanos
, Masculino
, Pessoa de Meia-Idade
, Análise Multivariada
RESUMO
BACKGROUND: Dictyostelium discoideum is a eukaryote with a simple lifestyle and a relatively small genome whose sequence has been fully determined. It is widely used for studies on cell signaling, movement and multicellular development. Ras guanine-nucleotide exchange factors (RasGEFs) are the proteins that activate Ras and thus lie near the top of many signaling pathways. They are particularly important for signaling in development and chemotaxis in many organisms, including Dictyostelium. RESULTS: We have searched the genome for sequences encoding RasGEFs. Despite its relative simplicity, we find that the Dictyostelium genome encodes at least 25 RasGEFs, with a few other genes encoding only parts of the RasGEF consensus domains. All appear to be expressed at some point in development. The 25 genes include a wide variety of domain structures, most of which have not been seen in other organisms. The LisH domain, which is associated with microtubule binding, is seen particularly frequently; other domains that confer interactions with the cytoskeleton are also common. Disruption of a sample of the novel genes reveals that many have clear phenotypes, including altered morphology and defects in chemotaxis, slug phototaxis and thermotaxis. CONCLUSION: These results suggest that the unexpectedly large number of RasGEF genes reflects an evolutionary expansion of the range of Ras signaling rather than functional redundancy or the presence of multiple pseudogenes.
Assuntos
Dictyostelium/genética , Genes de Protozoários/genética , Genoma de Protozoário/genética , Fatores ras de Troca de Nucleotídeo Guanina/genética , Fatores ras de Troca de Nucleotídeo Guanina/metabolismo , Actinas/genética , Animais , Dictyostelium/citologia , Regulação da Expressão Gênica no Desenvolvimento , Vida Livre de Germes , Movimento , Mutação/genética , Fenótipo , Filogenia , Ligação Proteica , Estrutura Terciária de Proteína , Análise de Sequência de DNA , Fatores ras de Troca de Nucleotídeo Guanina/química , Proteínas rho de Ligação ao GTP/metabolismoRESUMO
Rhobtb2 is a candidate tumor suppressor located on human chromosome 8p21, a region commonly deleted in cancer. Rhobtb2 is homozygously deleted in 3.5% of primary breast cancers, and gene expression is ablated in approximately 50% of breast and lung cancer cell lines. RhoBTB2 is an 83-kD, atypical Rho GTPase of unknown function, comprising an N-terminal Rho GTPase domain and two tandem BTB domains. In this report, we demonstrate that RhoBTB2 binds to the ubiquitin ligase scaffold, Cul3, via its first BTB domain and show in vitro and in vivo that RhoBTB2 is a substrate for a Cul3-based ubiquitin ligase complex. Moreover, we show that a RhoBTB2 missense mutant identified in a lung cancer cell line is neither able to bind Cul3 nor is it regulated by the ubiquitin/proteasome system, resulting in increased RhoBTB2 protein levels in vivo. We suggest a model in which RhoBTB2 functions as a tumor suppressor by recruiting proteins to a Cul3 ubiquitin ligase complex for degradation.
Assuntos
Ubiquitina-Proteína Ligases/metabolismo , Proteínas rho de Ligação ao GTP/metabolismo , Animais , Regulação Neoplásica da Expressão Gênica/fisiologia , Humanos , Neoplasias Pulmonares/metabolismo , Camundongos , Ligação Proteica , Estrutura Terciária de Proteína , Técnicas do Sistema de Duplo-HíbridoRESUMO
Loss of either of the Ras pathway members RasS or GefB causes growing Dictyostelium cells to move aberrantly rapidly. In this study, we describe the changes in motility that underlie these phenotypes using computer-assisted 3D dynamic image analysis. Unexpectedly, the two mutants use different mechanisms to achieve rapid migration. The rasS(-) cells' motility is characterised by highly dynamic cell morphology, with rapidly extending and retracting pseudopodia. The gefB(-) cells do not have an unusually dynamic morphology, and achieve their efficient translocation by the continual remodelling of an existing dominant anterior pseudopodium. In spite of these dramatic changes in pseudopodium behaviour, the underlying motility cycle of both mutants remains normal. The levels of F-actin in both mutant cell lines are significantly elevated with respect to the wild-type parental cells, suggesting a possible biochemical basis for these emphatic phenotypes.