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1.
PLoS Pathog ; 20(6): e1012252, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38833496

RESUMEN

Microbial pathogenicity often depends on the route of infection. For instance, P. aeruginosa or S. marcescens cause acute systemic infections when low numbers of bacteria are injected into D. melanogaster flies whereas flies succumb much slower to the continuous ingestion of these pathogens, even though both manage to escape from the gut compartment and reach the hemocoel. Here, we have developed a latent P. aeruginosa infection model by feeding flies on the bacteria for a short period. The bacteria stably colonize internal tissues yet hardly cause any damage since latently-infected flies live almost as long as noninfected control flies. The apparently dormant bacteria display particular characteristics in terms of bacterial colony morphology, composition of the outer cell wall, and motility. The virulence of these bacteria can however be reactivated upon wounding the host. We show that melanization but not the cellular or the systemic humoral response is the predominant host defense that establishes latency and may coerce the bacteria to a dormant state. In addition, the lasting activation of the melanization responses in latently-infected flies provides a degree of protection to the host against a secondary fungal infection. Latent infection by an ingested pathogen protects against a variety of homologous or heterologous systemic secondary infectious challenges, a situation previously described for the endosymbiotic Wolbachia bacteria, a guard against viral infections.


Asunto(s)
Drosophila melanogaster , Inmunidad Innata , Infecciones por Pseudomonas , Pseudomonas aeruginosa , Animales , Drosophila melanogaster/microbiología , Drosophila melanogaster/inmunología , Pseudomonas aeruginosa/patogenicidad , Pseudomonas aeruginosa/inmunología , Infecciones por Pseudomonas/inmunología , Infecciones por Pseudomonas/microbiología , Virulencia , Modelos Animales de Enfermedad , Interacciones Huésped-Patógeno/inmunología
2.
Proc Natl Acad Sci U S A ; 120(12): e2205140120, 2023 03 21.
Artículo en Inglés | MEDLINE | ID: mdl-36917667

RESUMEN

The Drosophila systemic immune response against many Gram-positive bacteria and fungi is mediated by the Toll pathway. How Toll-regulated effectors actually fulfill this role remains poorly understood as the known Toll-regulated antimicrobial peptide (AMP) genes are active only against filamentous fungi and not against Gram-positive bacteria or yeasts. Besides AMPs, two families of peptides secreted in response to infectious stimuli that activate the Toll pathway have been identified, namely Bomanins and peptides derived from a polyprotein precursor known as Baramicin A (BaraA). Unexpectedly, the deletion of a cluster of 10 Bomanins phenocopies the Toll mutant phenotype of susceptibility to infections. Here, we demonstrate that BaraA is required specifically in the host defense against Enterococcus faecalis and against the entomopathogenic fungus Metarhizium robertsii, albeit the fungal burden is not altered in BaraA mutants. BaraA protects the fly from the action of distinct toxins secreted by these Gram-positive and fungal pathogens, respectively, Enterocin V and Destruxin A. The injection of Destruxin A leads to the rapid paralysis of flies, whether wild type (WT) or mutant. However, a larger fraction of wild-type than BaraA flies recovers from paralysis within 5 to 10 h. BaraAs' function in protecting the host from the deleterious action of Destruxin is required in glial cells, highlighting a resilience role for the Toll pathway in the nervous system against microbial virulence factors. Thus, in complement to the current paradigm, innate immunity can cope effectively with the effects of toxins secreted by pathogens through the secretion of dedicated peptides, independently of xenobiotics detoxification pathways.


Asunto(s)
Proteínas de Drosophila , Drosophila , Animales , Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Proteínas de Drosophila/metabolismo , Receptores Toll-Like/metabolismo , Transducción de Señal , Péptidos/metabolismo , Hongos/metabolismo , Bacterias Grampositivas/metabolismo
3.
EMBO Rep ; 24(1): e56036, 2023 01 09.
Artículo en Inglés | MEDLINE | ID: mdl-36322050

RESUMEN

Host defense against infections encompasses both resistance, which targets microorganisms for neutralization or elimination, and resilience/disease tolerance, which allows the host to withstand/tolerate pathogens and repair damages. In Drosophila, the Toll signaling pathway is thought to mediate resistance against fungal infections by regulating the secretion of antimicrobial peptides, potentially including Bomanins. We find that Aspergillus fumigatus kills Drosophila Toll pathway mutants without invasion because its dissemination is blocked by melanization, suggesting a role for Toll in host defense distinct from resistance. We report that mutants affecting the Toll pathway or the 55C Bomanin locus are susceptible to the injection of two Aspergillus mycotoxins, restrictocin and verruculogen. The vulnerability of 55C deletion mutants to these mycotoxins is rescued by the overexpression of Bomanins specific to each challenge. Mechanistically, flies in which BomS6 is expressed in the nervous system exhibit an enhanced recovery from the tremors induced by injected verruculogen and display improved survival. Thus, innate immunity also protects the host against the action of microbial toxins through secreted peptides and thereby increases its resilience to infection.


Asunto(s)
Proteínas de Drosophila , Micotoxinas , Animales , Drosophila/genética , Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Proteínas de Drosophila/metabolismo , Micotoxinas/metabolismo , Aspergillus/genética , Aspergillus/metabolismo , Inmunidad Innata
4.
Cell ; 140(1): 148-60, 2010 Jan 08.
Artículo en Inglés | MEDLINE | ID: mdl-20074523

RESUMEN

Over 1 billion people are estimated to be overweight, placing them at risk for diabetes, cardiovascular disease, and cancer. We performed a systems-level genetic dissection of adiposity regulation using genome-wide RNAi screening in adult Drosophila. As a follow-up, the resulting approximately 500 candidate obesity genes were functionally classified using muscle-, oenocyte-, fat-body-, and neuronal-specific knockdown in vivo and revealed hedgehog signaling as the top-scoring fat-body-specific pathway. To extrapolate these findings into mammals, we generated fat-specific hedgehog-activation mutant mice. Intriguingly, these mice displayed near total loss of white, but not brown, fat compartments. Mechanistically, activation of hedgehog signaling irreversibly blocked differentiation of white adipocytes through direct, coordinate modulation of early adipogenic factors. These findings identify a role for hedgehog signaling in white/brown adipocyte determination and link in vivo RNAi-based scanning of the Drosophila genome to regulation of adipocyte cell fate in mammals.


Asunto(s)
Proteínas de Drosophila/metabolismo , Proteínas Hedgehog/metabolismo , Obesidad/genética , Adipocitos Marrones/metabolismo , Adipocitos Blancos/metabolismo , Adipogénesis , Animales , AMP Cíclico/metabolismo , Glucocorticoides/metabolismo , Humanos , Ratones , Ratones Noqueados , Células Musculares/metabolismo , Proteínas Represoras/genética
5.
Med Mycol ; 61(4)2023 Apr 03.
Artículo en Inglés | MEDLINE | ID: mdl-36931899

RESUMEN

Genome-wide comparisons have shown Talaromyces marneffei possessed a stable mating type locus in its meiosis genes. But the function of the mating type locus in T. marneffei is not clear. The potential sex recombination might lead to problems in clinical, such as the evolution of increased resistance to antifungal drugs and virulence. To determine the mating type in a sample of 107 T. marneffei isolates and to explore the possible relationship between fungus virulence and mating type or source. We used PCR analysis to determine the distribution of mating type genes and also analyzed the relationship between mating type and isolated sources (including HIV-positive patients, HIV-negative patients, bamboo rats, and the environment). Further, the Drosophila melanogaster model of infection was used to compare the differences of virulence in mating type and sources. Our results showed the entire sample population of T. marneffei with an overabundance of MAT1-2 alleles, but with a higher ratio of MAT1-1 in the isolates from HIV-negative patients. However, no significant differences in the survival of the D. melanogaster infected neither with MAT1-1 (6.5 days) nor MAT1-2 (4 days) isolates. Similar results were also observed in virulence analysis tested with different sources of isolates. So, we found that all isolates bore single mating type idiomorphs and unequal distribution. The distribution of the MAT genes seems related to different sources. And the virulence differences are independent of mating type genotype and source.


Our work shows the entire sample population of 107 Talaromyces marneffei isolates with an overabundance of MAT1-2 alleles, but with a higher ratio of MAT1-1 in the isolates from HIV-negative patients. And fungus virulence is independent of mating type genotype and source in the Drosophila melanogaster model.


Asunto(s)
Infecciones por VIH , Talaromyces , Animales , Drosophila melanogaster , Genes del Tipo Sexual de los Hongos , Talaromyces/genética , Infecciones por VIH/veterinaria
6.
Immunogenetics ; 74(1): 35-62, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-35092465

RESUMEN

Insects occupy a central position in the biosphere. They are able to resist infections even though they lack an adaptive immune system. Drosophila melanogaster has been used as a potent genetic model to understand innate immunity both in invertebrates and vertebrates. Its immune system includes both humoral and cellular arms. Here, we review how the distinct immune responses are triggered upon sensing infections, with an emphasis on the mechanisms that lead to systemic humoral immune responses. As in plants, the components of the cell wall of microorganisms are detected by dedicated receptors. There is also an induction of the systemic immune response upon sensing the proteolytic activities of microbial virulence factors. The antiviral response mostly relies on sensing double-stranded RNAs generated during the viral infection cycle. This event subsequently triggers either the viral short interfering RNA pathway or a cGAS-like/STING/NF-κB signaling pathway.


Asunto(s)
Drosophila melanogaster , Modelos Genéticos , Animales , Drosophila melanogaster/genética , Inmunidad Innata/genética , FN-kappa B/genética , FN-kappa B/metabolismo , Transducción de Señal/genética
7.
EMBO Rep ; 19(5)2018 05.
Artículo en Inglés | MEDLINE | ID: mdl-29523648

RESUMEN

When Drosophila melanogaster feeds on Pseudomonas aeruginosa, some bacteria cross the intestinal barrier and eventually proliferate in the hemocoel. This process is limited by hemocytes through phagocytosis. P. aeruginosa requires the quorum-sensing regulator RhlR to elude the cellular immune response of the fly. RhlI synthesizes the autoinducer signal that activates RhlR. Here, we show that rhlI mutants are unexpectedly more virulent than rhlR mutants, both in fly and in nematode intestinal infection models, suggesting that RhlR has RhlI-independent functions. We also report that RhlR protects P. aeruginosa from opsonization mediated by the Drosophila thioester-containing protein 4 (Tep4). RhlR mutant bacteria show higher levels of Tep4-mediated opsonization, as compared to rhlI mutants, which prevents lethal bacteremia in the Drosophila hemocoel. In contrast, in a septic model of infection, in which bacteria are introduced directly into the hemocoel, Tep4 mutant flies are more resistant to wild-type P. aeruginosa, but not to the rhlR mutant. Thus, depending on the infection route, the Tep4 opsonin can either be protective or detrimental to host defense.


Asunto(s)
Proteínas Bacterianas/genética , ARN Helicasas DEAD-box/genética , Ligasas/genética , Fagocitosis , Pseudomonas aeruginosa/genética , Percepción de Quorum/genética , Factores de Transcripción/genética , Animales , Caenorhabditis elegans/microbiología , Proteínas de Drosophila/genética , Proteínas de Drosophila/inmunología , Drosophila melanogaster/inmunología , Drosophila melanogaster/microbiología , Regulación Bacteriana de la Expresión Génica , Intestinos/inmunología , Intestinos/microbiología , Pseudomonas aeruginosa/patogenicidad , Receptores de Reconocimiento de Patrones/inmunología , Virulencia
8.
Bioorg Med Chem Lett ; 28(15): 2631-2635, 2018 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-29945796

RESUMEN

Fipronil is a phenyl pyrazole molecule widely used across the world as both insecticide and veterinary drug. The main goal of this work was to synthesize a fluorescently labeled fipronil derivative for cellular imaging without affecting its intrinsic properties. We selected fluorescein as fluorescent probe and we investigated different strategies for stable chemical ligation between both entities, such as thiourea and direct peptide bond. While thiourea bond displayed low stability, direct peptide bond was difficult to achieve due to problems of steric hindrance. The best result was obtained by conjugation using click chemistry, which allowed to obtain fipronil stably labeled with the fluorescent probe.


Asunto(s)
Antiparasitarios/química , Antiparasitarios/síntesis química , Fluoresceína/química , Colorantes Fluorescentes/química , Insecticidas/química , Insecticidas/síntesis química , Pirazoles/química , Pirazoles/síntesis química , Amidas/química , Antiparasitarios/toxicidad , Química Clic , Estabilidad de Medicamentos , Insecticidas/toxicidad , Pirazoles/toxicidad , Tiourea/química , Drogas Veterinarias
9.
PLoS Pathog ; 10(6): e1004211, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24945925

RESUMEN

The opportunistic fungal pathogen Candida glabrata is a frequent cause of candidiasis, causing infections ranging from superficial to life-threatening disseminated disease. The inherent tolerance of C. glabrata to azole drugs makes this pathogen a serious clinical threat. To identify novel genes implicated in antifungal drug tolerance, we have constructed a large-scale C. glabrata deletion library consisting of 619 unique, individually bar-coded mutant strains, each lacking one specific gene, all together representing almost 12% of the genome. Functional analysis of this library in a series of phenotypic and fitness assays identified numerous genes required for growth of C. glabrata under normal or specific stress conditions, as well as a number of novel genes involved in tolerance to clinically important antifungal drugs such as azoles and echinocandins. We identified 38 deletion strains displaying strongly increased susceptibility to caspofungin, 28 of which encoding proteins that have not previously been linked to echinocandin tolerance. Our results demonstrate the potential of the C. glabrata mutant collection as a valuable resource in functional genomics studies of this important fungal pathogen of humans, and to facilitate the identification of putative novel antifungal drug target and virulence genes.


Asunto(s)
Antifúngicos/farmacología , Azoles/farmacología , Candida glabrata/genética , Farmacorresistencia Fúngica/genética , Equinocandinas/farmacología , Biopelículas/crecimiento & desarrollo , Candida glabrata/crecimiento & desarrollo , Candidiasis/tratamiento farmacológico , Caspofungina , Pared Celular/efectos de los fármacos , Pared Celular/genética , Proteínas Fúngicas/genética , Eliminación de Gen , Técnicas de Inactivación de Genes , Biblioteca de Genes , Lipopéptidos , Pruebas de Sensibilidad Microbiana , Presión Osmótica , Fenotipo
10.
J Immunol ; 190(6): 2818-27, 2013 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-23401590

RESUMEN

The pathogenicity of Candida glabrata to patients remains poorly understood for lack of convenient animal models to screen large numbers of mutants for altered virulence. In this study, we explore the minihost model Drosophila melanogaster from the dual perspective of host and pathogen. As in vertebrates, wild-type flies contain C. glabrata systemic infections yet are unable to kill the injected yeasts. As for other fungal infections in Drosophila, the Toll pathway restrains C. glabrata proliferation. Persistent C. glabrata yeasts in wild-type flies do not appear to be able to take shelter in hemocytes from the action of the Toll pathway, the effectors of which remain to be identified. Toll pathway mutant flies succumb to injected C. glabrata. In this immunosuppressed background, cellular defenses provide a residual level of protection. Although both the Gram-negative binding protein 3 pattern recognition receptor and the Persephone protease-dependent detection pathway are required for Toll pathway activation by C. glabrata, only GNBP3, and not psh mutants, are susceptible to the infection. Both Candida albicans and C. glabrata are restrained by the Toll pathway, yet the comparative study of phenoloxidase activation reveals a differential activity of the Toll pathway against these two fungal pathogens. Finally, we establish that the high-osmolarity glycerol pathway and yapsins are required for virulence of C. glabrata in this model. Unexpectedly, yapsins do not appear to be required to counteract the cellular immune response but are needed for the colonization of the wild-type host.


Asunto(s)
Candida glabrata/patogenicidad , Candidiasis/inmunología , Candidiasis/microbiología , Proteínas de Drosophila/fisiología , Drosophila melanogaster/inmunología , Transducción de Señal/inmunología , Receptores Toll-Like/fisiología , Proteínas Adaptadoras Transductoras de Señales/deficiencia , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Antígenos de Diferenciación/genética , Candida glabrata/inmunología , Candidiasis/genética , Células Cultivadas , Modelos Animales de Enfermedad , Proteínas de Drosophila/deficiencia , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Fagocitosis/genética , Fagocitosis/inmunología , Receptores Inmunológicos/deficiencia , Receptores Inmunológicos/genética , Transducción de Señal/genética , Receptores Toll-Like/deficiencia , Receptores Toll-Like/genética , Virulencia/genética , Virulencia/inmunología
11.
Proc Natl Acad Sci U S A ; 108(42): 17378-83, 2011 Oct 18.
Artículo en Inglés | MEDLINE | ID: mdl-21987808

RESUMEN

An in-depth mechanistic understanding of microbial infection necessitates a molecular dissection of host-pathogen relationships. Both Drosophila melanogaster and Pseudomonas aeruginosa have been intensively studied. Here, we analyze the infection of D. melanogaster by P. aeruginosa by using mutants in both host and pathogen. We show that orally ingested P. aeruginosa crosses the intestinal barrier and then proliferates in the hemolymph, thereby causing the infected flies to die of bacteremia. Host defenses against ingested P. aeruginosa included an immune deficiency (IMD) response in the intestinal epithelium, systemic Toll and IMD pathway responses, and a cellular immune response controlling bacteria in the hemocoel. Although the observed cellular and intestinal immune responses appeared to act throughout the course of the infection, there was a late onset of the systemic IMD and Toll responses. In this oral infection model, P. aeruginosa PA14 did not require its type III secretion system or other well-studied virulence factors such as the two-component response regulator GacA or the protease AprA for virulence. In contrast, the quorum-sensing transcription factor RhlR, but surprisingly not LasR, played a key role in counteracting the cellular immune response against PA14, possibly at an early stage when only a few bacteria are present in the hemocoel. These results illustrate the power of studying infection from the dual perspective of host and pathogen by revealing that RhlR plays a more complex role during pathogenesis than previously appreciated.


Asunto(s)
Proteínas Bacterianas/inmunología , Drosophila melanogaster/inmunología , Drosophila melanogaster/microbiología , Inmunidad Celular , Pseudomonas aeruginosa/inmunología , Pseudomonas aeruginosa/patogenicidad , Administración Oral , Animales , Animales Modificados Genéticamente , Bacteriemia/inmunología , Proteínas Bacterianas/genética , Modelos Animales de Enfermedad , Drosophila melanogaster/genética , Genes de Insecto , Genes Virales , Hemolinfa/microbiología , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Mutación , Infecciones por Pseudomonas/inmunología , Pseudomonas aeruginosa/genética , Percepción de Quorum/inmunología , Transactivadores/genética , Transactivadores/inmunología , Virulencia/inmunología
12.
Fly (Austin) ; 18(1): 2398300, 2024 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-39239739

RESUMEN

Talaromycosis, caused by Talaromyces marneffei (T. marneffei, formerly known as Penicillium marneffei), is an opportunistic invasive mycosis endemic in tropical and subtropical areas of Asia with high mortality rate. Despite various infection models established to study the immunological interaction between T. marneffei and the host, the pathogenicity of this fungus is not yet fully understood. So far, Drosophila melanogaster, a well-established genetic model organism to study innate immunity, has not been used in related research on T. marneffei. In this study, we provide the initial characterization of a systemic infection model of T. marneffei in the D. melanogaster host. Survival curves and fungal loads were tested as well as Toll pathway activation was quantified by RT-qPCR of several antimicrobial peptide (AMP) genes including Drosomycin, Metchnikowin, and Bomanin Short 1. We discovered that whereas most wild-type flies were able to overcome the infection, MyD88 or Toll mutant flies failed to prevent fungal dissemination and proliferation and ultimately succumbed to this challenge. Unexpectedly, the induction of classical Toll pathway activation readouts, Drosomycin and Bomanin Short 1, by live or killed T. marneffei was quite limited in wild-type flies, suggesting that the fungus largely escapes detection by the systemic immune system. This unusual situation of a poor systemic activation of the Toll pathway and a strong susceptibility phenotype of MyD88/Toll might be accounted for by a requirement for this host defence in only specific tissues, a hypothesis that remains to be rigorously tested.


Asunto(s)
Proteínas de Drosophila , Drosophila melanogaster , Factor 88 de Diferenciación Mieloide , Talaromyces , Receptores Toll-Like , Animales , Talaromyces/genética , Talaromyces/metabolismo , Factor 88 de Diferenciación Mieloide/metabolismo , Factor 88 de Diferenciación Mieloide/genética , Drosophila melanogaster/microbiología , Drosophila melanogaster/inmunología , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Receptores Toll-Like/metabolismo , Receptores Toll-Like/genética , Micosis/inmunología , Micosis/microbiología , Inmunidad Innata , Transducción de Señal , Antígenos de Diferenciación , Receptores Inmunológicos , Proteínas Adaptadoras Transductoras de Señales
13.
Int Immunopharmacol ; 129: 111559, 2024 Mar 10.
Artículo en Inglés | MEDLINE | ID: mdl-38330794

RESUMEN

Antibiotic-resistant Serratia marcescens (Sm) is known to cause bloodstream infections, pneumonia, etc. The nod-like receptor family, pyrin domain-containing 3 (NLRP3), has been implicated in various lung infections. Yet, its role in Sm-induced pneumonia was not well understood. In our study, we discovered that deletion of Nlrp3 in mice significantly improved Sm-induced survival rates, reduced bacterial loads in the lungs, bronchoalveolar lavage fluid (BALF), and bloodstream, and mitigated the severity of acute lung injury (ALI) compared to wild-type (WT) mice. Mechanistically, we observed that 24 h post-Sm infection, NLRP3 inflammasome activation occurred, leading to gasdermin D NH2-terminal (GSDMD-NT)-induced pyroptosis in macrophages and IL-1ß secretion. The NLRP3 or NLRP3 inflammasome influenced the expression PD-L1 and PD-1, as well as the count of PD-L1 or PD-1-expressing macrophages, alveolar macrophages, interstitial macrophages, PD-L1-expressing neutrophils, and the count of macrophage receptors with collagenous structure (MARCO)-expressing macrophages, particularly MARCO+ alveolar macrophages. The frequency of MARCO+ alveolar macrophages, PD-1 expression, particularly PD-1+ interstitial macrophages were negatively or positively correlated with the Sm load, respectively. Additionally, IL-1ß levels in BALF correlated with three features of acute lung injury: histologic score, protein concentration and neutrophil count in BALF. Consequently, our findings suggest that Nlrp3 deletion offers protection agaisnt acute Sm pneumonia in mice by inhibiting inflammasome activation and reducing Sm infection-induced PD-L1/PD-1 or MARCO expression, particularly in macrophages. This highlights potential therapeutic targets for Sm and other gram-negative bacteria-induced acute pneumonia.


Asunto(s)
Lesión Pulmonar Aguda , Neumonía , Ratones , Animales , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Inflamasomas/metabolismo , Receptor de Muerte Celular Programada 1/metabolismo , Serratia marcescens/genética , Serratia marcescens/metabolismo , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Neumonía/metabolismo , Macrófagos/metabolismo , Lesión Pulmonar Aguda/inducido químicamente , Interleucina-1beta/metabolismo , Lipopolisacáridos/farmacología , Ratones Noqueados
14.
Nat Commun ; 15(1): 5713, 2024 Jul 08.
Artículo en Inglés | MEDLINE | ID: mdl-38977661

RESUMEN

Cellular senescence is characterized by a decrease in protein synthesis, although the underlying processes are mostly unclear. Chemical modifications to transfer RNAs (tRNAs) frequently influence tRNA activity, which is crucial for translation. We describe how tRNA N7-methylguanosine (m7G46) methylation, catalyzed by METTL1-WDR4, regulates translation and influences senescence phenotypes. Mettl1/Wdr4 and m7G gradually diminish with senescence and aging. A decrease in METTL1 causes a reduction in tRNAs, especially those with the m7G modification, via the rapid tRNA degradation (RTD) pathway. The decreases cause ribosomes to stall at certain codons, impeding the translation of mRNA that is essential in pathways such as Wnt signaling and ribosome biogenesis. Furthermore, chronic ribosome stalling stimulates the ribotoxic and integrative stress responses, which induce senescence-associated secretory phenotype. Moreover, restoring eEF1A protein mitigates senescence phenotypes caused by METTL1 deficiency by reducing RTD. Our findings demonstrate that tRNA m7G modification is essential for preventing premature senescence and aging by enabling efficient mRNA translation.


Asunto(s)
Senescencia Celular , Guanosina , Metiltransferasas , Biosíntesis de Proteínas , ARN de Transferencia , Senescencia Celular/genética , ARN de Transferencia/metabolismo , ARN de Transferencia/genética , Metiltransferasas/metabolismo , Metiltransferasas/genética , Guanosina/análogos & derivados , Guanosina/metabolismo , Metilación , Humanos , Ribosomas/metabolismo , Envejecimiento/metabolismo , Envejecimiento/genética , ARN Mensajero/metabolismo , ARN Mensajero/genética , Animales , Factor 1 de Elongación Peptídica/metabolismo , Factor 1 de Elongación Peptídica/genética , Estabilidad del ARN
15.
iScience ; 26(9): 107490, 2023 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-37636057

RESUMEN

Upon exposure to a bacterial pore-forming toxin, enterocytes rapidly purge their apical cytoplasm into the gut lumen, resulting in a thin intestinal epithelium. The enterocytes regain their original shape and thickness within 16 h after the ingestion of the bacteria. Here, we show that the regrowth of Drosophila enterocytes entails an inversion of metabolic fluxes from the organism back toward the intestine. We identify a proton-assisted transporter, Arcus, that is required for the reverse absorption of amino acids and the timely recovery of the intestinal epithelium. Arcus is required for a peak of amino acids appearing in the hemolymph shortly after infection. The regrowth of enterocytes involves the insulin signaling pathway and Myc. The purge decreases Myc mRNA levels, which subsequently remain at low levels in the arcus mutant. Interestingly, the action of arcus and Myc in the intestinal epithelium is not cell-autonomous, suggesting amino acid fluxes within the intestinal epithelium.

16.
Res Microbiol ; 174(6): 104089, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37348743

RESUMEN

Bacillus thuringiensis israelensis is largely regarded as the most selective, safe and ecofriendly biopesticide used for the control of insect vectors of human diseases. Bti enthomopathogenicity relies on the Cry and Cyt δ-endotoxins, produced as crystalline inclusions during sporulation. Insecticidal selectivity of Bti is mainly ascribed to the binding of the Cry toxins to receptors in the gut of target insects. However, the contribution of epithelial defenses in limiting Bti side effects in non-target species remains largely unexplored. Here, taking advantage of the genetically tractable Drosophila melanogaster model and its amenability for deciphering highly conserved innate immune defenses, we unravel a central role of the NF-κB factor Relish in the protection against the effects of ingested Bti spores in a non-susceptible host. Intriguingly, our data indicate that the Bti-induced Relish response is independent of its canonical activation downstream of peptidoglycan sensing and does not involve its longstanding role in the regulation of antimicrobial peptides encoding genes. In contrast, our data highlight a novel enterocyte specific function of Relish that is essential for preventing general septicemia following Bti oral infections strictly when producing δ-endotoxins. Altogether, our data provide novel insights into Bti-hosts interactions of prominent interest for the optimization and sustainability of insects' biocontrol strategies.


Asunto(s)
Bacillus thuringiensis , Endotoxinas , Animales , Humanos , Endotoxinas/genética , Endotoxinas/metabolismo , Endotoxinas/farmacología , Bacillus thuringiensis/genética , FN-kappa B/metabolismo , Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Toxinas de Bacillus thuringiensis/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Hemolisinas/metabolismo , Proteínas Hemolisinas/farmacología
17.
Front Immunol ; 13: 858360, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35493511

RESUMEN

Microsporidia are obligate intracellular parasites able to infest specifically a large range of species, including insects. The knowledge about the biology of microsporidial infections remains confined to mostly descriptive studies, including molecular approaches such as transcriptomics or proteomics. Thus, functional data to understand insect host defenses are currently lacking. Here, we have undertaken a genetic analysis of known host defenses of the Drosophila melanogaster using an infection model whereby Tubulinosema ratisbonensis spores are directly injected in this insect. We find that phagocytosis does confer some protection in this infection model. In contrast, the systemic immune response, extracellular reactive oxygen species, thioester proteins, xenophagy, and intracellular antiviral response pathways do not appear to be involved in the resistance against this parasite. Unexpectedly, several genes such as PGRP-LE seem to promote this infection. The prophenol oxidases that mediate melanization have different functions; PPO1 presents a phenotype similar to that of PGRP-LE whereas that of PPO2 suggests a function in the resilience to infection. Similarly, eiger and Unpaired3, which encode two cytokines secreted by hemocytes display a resilience phenotype with a strong susceptibility to T. ratisbonensis.


Asunto(s)
Drosophila melanogaster , Microsporidiosis , Animales , Hemocitos , Inmunidad , Fagocitosis
18.
Sci Rep ; 12(1): 3068, 2022 02 23.
Artículo en Inglés | MEDLINE | ID: mdl-35197500

RESUMEN

Serratia marcescens is an opportunistic bacterium that infects a wide range of hosts including humans. It is a potent pathogen in a septic injury model of Drosophila melanogaster since a few bacteria directly injected in the body cavity kill the insect within a day. In contrast, flies do not succumb to ingested bacteria for days even though some bacteria cross the intestinal barrier into the hemolymph within hours. The mechanisms by which S. marcescens attacks enterocytes and damages the intestinal epithelium remain uncharacterized. To better understand intestinal infections, we performed a genetic screen for loss of virulence of ingested S. marcescens and identified FliR, a structural component of the flagellum, as a virulence factor. Next, we compared the virulence of two flagellum mutants fliR and flhD in two distinct S. marcescens strains. Both genes are required for S. marcescens to escape the gut lumen into the hemocoel, indicating that the flagellum plays an important role for the passage of bacteria through the intestinal barrier. Unexpectedly, fliR but not flhD is involved in S. marcescens-mediated damages of the intestinal epithelium that ultimately contribute to the demise of the host. Our results therefore suggest a flagellum-independent role for fliR in bacterial virulence.


Asunto(s)
Proteínas Bacterianas/genética , Proteínas Bacterianas/fisiología , Drosophila melanogaster/microbiología , Flagelos/genética , Flagelos/fisiología , Gastroenteritis/microbiología , Mucosa Intestinal/microbiología , Proteínas de la Membrana/genética , Proteínas de la Membrana/fisiología , Infecciones por Serratia , Serratia marcescens/genética , Serratia marcescens/patogenicidad , Animales , Modelos Animales de Enfermedad , Mucosa Intestinal/patología , Mutación , Virulencia/genética
19.
Sci Signal ; 15(729): eabm5011, 2022 04 12.
Artículo en Inglés | MEDLINE | ID: mdl-35412849

RESUMEN

Toll-like receptor (TLR) stimulation induces glycolysis and the production of mitochondrial reactive oxygen species (ROS), both of which are critical for inflammatory responses in macrophages. Here, we demonstrated that cyclin J, a TLR-inducible member of the cyclin family, reduced cytokine production in macrophages by coordinately controlling glycolysis and mitochondrial functions. Cyclin J interacted with cyclin-dependent kinases (CDKs), which increased the phosphorylation of a subset of CDK substrates, including the transcription factor FoxK1 and the GTPase Drp1. Cyclin J-dependent phosphorylation of FoxK1 decreased the transcription of glycolytic genes and Hif-1α activation, whereas hyperactivation of Drp1 by cyclin J-dependent phosphorylation promoted mitochondrial fragmentation and impaired the production of mitochondrial ROS. In mice, cyclin J in macrophages limited the growth of tumor xenografts and protected against LPS-induced shock but increased the susceptibility to bacterial infection. Collectively, our findings indicate that cyclin J-CDK signaling promotes antitumor immunity and the resolution of inflammation by opposing the metabolic changes that drive inflammatory responses in macrophages.


Asunto(s)
Inmunidad Innata , Macrófagos , Animales , Quinasas Ciclina-Dependientes/genética , Quinasas Ciclina-Dependientes/metabolismo , Ciclinas/metabolismo , Factores de Transcripción Forkhead/metabolismo , Humanos , Macrófagos/metabolismo , Ratones , Especies Reactivas de Oxígeno/metabolismo , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismo
20.
Eur J Immunol ; 40(5): 1244-54, 2010 May.
Artículo en Inglés | MEDLINE | ID: mdl-20201042

RESUMEN

The Drosophila Toll-signaling pathway controls the systemic antifungal host response. Gram-negative binding protein 3 (GNBP3), a member of the beta-glucan recognition protein family senses fungal infections and activates this pathway. A second detection system perceives the activity of proteolytic fungal virulence factors and redundantly activates Toll. GNBP3(hades) mutant flies succumb more rapidly to Candida albicans and to entomopathogenic fungal infections than WT flies, despite normal triggering of the Toll pathway via the virulence detection system. These observations suggest that GNBP3 triggers antifungal defenses that are not dependent on activation of the Toll pathway. Here, we show that GNBP3 agglutinates fungal cells. Furthermore, it can activate melanization in a Toll-independent manner. Melanization is likely to be an essential defense against some fungal infections given that the entomopathogenic fungus Beauveria bassiana inhibits the activity of the main melanization enzymes, the phenol oxidases. Finally, we show that GNBP3 assembles "attack complexes", which comprise phenoloxidase and the necrotic serpin. We propose that Drosophila GNBP3 targets fungi immediately at the inception of the infection by bringing effector molecules in direct contact with the invading microorganisms.


Asunto(s)
Proteínas Portadoras/inmunología , Proteínas de Drosophila/inmunología , Drosophila melanogaster/inmunología , Hongos/inmunología , Melaninas/fisiología , Aglutinación , Animales , Beauveria/inmunología , Candida albicans/inmunología , Proteínas Portadoras/farmacología , Proteínas de Drosophila/farmacología , Proteínas de Drosophila/fisiología , Drosophila melanogaster/microbiología , Activación Enzimática , Hemolinfa/inmunología , Péptidos y Proteínas de Señalización Intracelular , Monofenol Monooxigenasa/fisiología , Complejos Multiproteicos/fisiología , Proteínas Recombinantes de Fusión/farmacología , Serpinas/fisiología , Esporas Fúngicas , Receptores Toll-Like/inmunología
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