RESUMO
Capicua (Cic) proteins are conserved HMG-box transcriptional repressors that control receptor tyrosine kinase (RTK) signaling responses and are implicated in human neurological syndromes and cancer. While Cic is known to exist as short (Cic-S) and long (Cic-L) isoforms with identical HMG-box and associated core regions but distinct N termini, most previous studies have focused on Cic-S, leaving the function of Cic-L unexplored. Here we show that Cic-L acts in two capacities during Drosophila oogenesis: 1) as a canonical sensor of RTK signaling in somatic follicle cells, and 2) as a regulator of postmitotic growth in germline nurse cells. In these latter cells, Cic-L behaves as a temporal signal that terminates endoreplicative growth before they dump their contents into the oocyte. We show that Cic-L is necessary and sufficient for nurse cell endoreplication arrest and induces both stabilization of CycE and down-regulation of Myc. Surprisingly, this function depends mainly on the Cic-L-specific N-terminal module, which is capable of acting independently of the Cic HMG-box-containing core. Mirroring these observations, basal metazoans possess truncated Cic-like proteins composed only of Cic-L N-terminal sequences, suggesting that this module plays unique, ancient roles unrelated to the canonical function of Cic.
Assuntos
Proteínas de Drosophila , Drosophila melanogaster , Proteínas HMGB , Oogênese , Proteínas Repressoras , Animais , Proteínas de Drosophila/genética , Proteínas de Drosophila/fisiologia , Drosophila melanogaster/fisiologia , Proteínas HMGB/genética , Proteínas HMGB/fisiologia , Oogênese/genética , Receptores Proteína Tirosina Quinases/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/fisiologiaRESUMO
CIC (also known as Capicua) is a transcriptional repressor negatively regulated by RAS/MAPK signaling. Whereas the functions of Cic have been well characterized in Drosophila, little is known about its role in mammals. CIC is inactivated in a variety of human tumors and has been implicated recently in the promotion of lung metastases. Here, we describe a mouse model in which we inactivated Cic by selectively disabling its DNA-binding activity, a mutation that causes derepression of its target genes. Germline Cic inactivation causes perinatal lethality due to lung differentiation defects. However, its systemic inactivation in adult mice induces T-cell acute lymphoblastic lymphoma (T-ALL), a tumor type known to carry CIC mutations, albeit with low incidence. Cic inactivation in mice induces T-ALL by a mechanism involving derepression of its well-known target, Etv4 Importantly, human T-ALL also relies on ETV4 expression for maintaining its oncogenic phenotype. Moreover, Cic inactivation renders T-ALL insensitive to MEK inhibitors in both mouse and human cell lines. Finally, we show that Ras-induced mouse T-ALL as well as human T-ALL carrying mutations in the RAS/MAPK pathway display a genetic signature indicative of Cic inactivation. These observations illustrate that CIC inactivation plays a key role in this human malignancy.
Assuntos
Leucemia-Linfoma Linfoblástico de Células T Precursoras/genética , Proteínas Repressoras/genética , Proteínas E1A de Adenovirus/metabolismo , Alelos , Animais , Neoplasias Encefálicas/genética , Linhagem Celular Tumoral , Desenvolvimento Embrionário/genética , Fibroblastos/metabolismo , Genes ras , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos , Mutação , Oligodendroglioma/genética , Leucemia-Linfoma Linfoblástico de Células T Precursoras/enzimologia , Leucemia-Linfoma Linfoblástico de Células T Precursoras/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-ets/genética , Transcrição GênicaRESUMO
The HMG-box protein Capicua (Cic) is a conserved transcriptional repressor that functions downstream of receptor tyrosine kinase (RTK) signaling pathways in a relatively simple switch: In the absence of signaling, Cic represses RTK-responsive genes by binding to nearly invariant sites in DNA, whereas activation of RTK signaling down-regulates Cic activity, leading to derepression of its targets. This mechanism controls gene expression in both Drosophila and mammals, but whether Cic can also function via other regulatory mechanisms remains unknown. Here, we characterize an RTK-independent role of Cic in regulating spatially restricted expression of Toll/IL-1 signaling targets in Drosophila embryogenesis. We show that Cic represses those targets by binding to suboptimal DNA sites of lower affinity than its known consensus sites. This binding depends on Dorsal/NF-κB, which translocates into the nucleus upon Toll activation and binds next to the Cic sites. As a result, Cic binds to and represses Toll targets only in regions with nuclear Dorsal. These results reveal a mode of Cic regulation unrelated to the well-established RTK/Cic depression axis and implicate cooperative binding in conjunction with low-affinity binding sites as an important mechanism of enhancer regulation. Given that Cic plays a role in many developmental and pathological processes in mammals, our results raise the possibility that some of these Cic functions are independent of RTK regulation and may depend on cofactor-assisted DNA binding.
Assuntos
Proteínas de Drosophila/metabolismo , Drosophila/genética , Proteínas HMGB/metabolismo , Receptores Proteína Tirosina Quinases/metabolismo , Proteínas Repressoras/metabolismo , Transdução de Sinais , Receptores Toll-Like/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Núcleo Celular/genética , Núcleo Celular/metabolismo , Drosophila/embriologia , Drosophila/enzimologia , Drosophila/metabolismo , Proteínas de Drosophila/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas HMGB/genética , Masculino , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Regiões Promotoras Genéticas , Receptores Proteína Tirosina Quinases/genética , Proteínas Repressoras/genética , Receptores Toll-Like/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
HMG-box proteins, including Sox/SRY (Sox) and TCF/LEF1 (TCF) family members, bind DNA via their HMG-box. This binding, however, is relatively weak and both Sox and TCF factors employ distinct mechanisms for enhancing their affinity and specificity for DNA. Here we report that Capicua (CIC), an HMG-box transcriptional repressor involved in Ras/MAPK signaling and cancer progression, employs an additional distinct mode of DNA binding that enables selective recognition of its targets. We find that, contrary to previous assumptions, the HMG-box of CIC does not bind DNA alone but instead requires a distant motif (referred to as C1) present at the C-terminus of all CIC proteins. The HMG-box and C1 domains are both necessary for binding specific TGAATGAA-like sites, do not function via dimerization, and are active in the absence of cofactors, suggesting that they form a bipartite structure for sequence-specific binding to DNA. We demonstrate that this binding mechanism operates throughout Drosophila development and in human cells, ensuring specific regulation of multiple CIC targets. It thus appears that HMG-box proteins generally depend on auxiliary DNA binding mechanisms for regulating their appropriate genomic targets, but that each sub-family has evolved unique strategies for this purpose. Finally, the key role of C1 in DNA binding also explains the fact that this domain is a hotspot for inactivating mutations in oligodendroglioma and other tumors, while being preserved in oncogenic CIC-DUX4 fusion chimeras associated to Ewing-like sarcomas.
Assuntos
DNA/genética , Proteínas de Drosophila/genética , Proteínas HMGB/genética , Proteínas de Grupo de Alta Mobilidade/genética , Mutação , Neoplasias/genética , Proteínas Repressoras/genética , Motivos de Aminoácidos/genética , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Sequência de Bases , Sítios de Ligação/genética , DNA/metabolismo , Drosophila/embriologia , Drosophila/genética , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Embrião não Mamífero/citologia , Embrião não Mamífero/embriologia , Embrião não Mamífero/metabolismo , Células HEK293 , Domínios HMG-Box/genética , Proteínas HMGB/metabolismo , Proteínas de Grupo de Alta Mobilidade/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Imuno-Histoquímica , Microscopia Confocal , Modelos Genéticos , Neoplasias/metabolismo , Ligação Proteica , Proteínas Repressoras/metabolismo , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
This article aims to describe the cutaneous manifestations observed in the Zika epidemic in Peru during 2016 and 2017, as well as discuss the potential differential diagnoses. During the outbreak, the main reason for seeking medical advice was the development of a pruriginous maculopapular rash with a marked papular component, which started on the chest and later generalized to the rest of the body. Similar manifestations were noted in adults, children, and pregnant women. Other manifestations such as conjunctivitis, edema, or petechiae on the palate were rare. We suggest that in areas that are endemic for arboviral infections, in the differential diagnosis of a rash one must consider infections such as dengue, Zika, or chikungunya viruses. In nonendemic areas, the diagnosis is more difficult, as the rash may result from other viral infections not transmitted by arthropods and/or reactive or inflammatory diseases (urticaria, atopic dermatitis, systemic lupus erythematosus). We recommend that primary care health personnel are trained in the recognition of the mucocutaneous lesions caused by Zika virus infection, which could contribute to the identification of suspicious cases, particularly pregnant women.
Assuntos
Exantema/etiologia , Pele/virologia , Infecção por Zika virus/epidemiologia , Zika virus , Diagnóstico Diferencial , Exantema/diagnóstico , Exantema/virologia , Humanos , Incidência , Peru/epidemiologia , Pele/patologia , Infecção por Zika virus/diagnóstico , Infecção por Zika virus/virologiaRESUMO
The transcriptional repressor Capicua (Cic) controls tissue patterning and restricts organ growth, and has been recently implicated in several cancers. Cic has emerged as a primary sensor of signaling downstream of the receptor tyrosine kinase (RTK)/extracellular signal-regulated kinase (ERK) pathway, but how Cic activity is regulated in different cellular contexts remains poorly understood. We found that the kinase Minibrain (Mnb, ortholog of mammalian DYRK1A), acting through the adaptor protein Wings apart (Wap), physically interacts with and phosphorylates the Cic protein. Mnb and Wap inhibit Cic function by limiting its transcriptional repressor activity. Down-regulation of Cic by Mnb/Wap is necessary for promoting the growth of multiple organs, including the wings, eyes, and the brain, and for proper tissue patterning in the wing. We have thus uncovered a previously unknown mechanism of down-regulation of Cic activity by Mnb and Wap, which operates independently from the ERK-mediated control of Cic. Therefore, Cic functions as an integrator of upstream signals that are essential for tissue patterning and organ growth. Finally, because DYRK1A and CIC exhibit, respectively, prooncogenic vs. tumor suppressor activities in human oligodendroglioma, our results raise the possibility that DYRK1A may also down-regulate CIC in human cells.
Assuntos
Padronização Corporal/genética , Proteínas de Drosophila/genética , Drosophila/genética , Proteínas HMGB/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas Tirosina Quinases/genética , Proteínas Repressoras/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Drosophila/crescimento & desenvolvimento , Proteínas de Drosophila/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Proteínas HMGB/biossíntese , Humanos , Neoplasias/genética , Fosforilação , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Repressoras/biossíntese , Asas de Animais/crescimento & desenvolvimento , Quinases DyrkRESUMO
Epithelial renewal in the Drosophila intestine is orchestrated by Intestinal Stem Cells (ISCs). Following damage or stress the intestinal epithelium produces ligands that activate the epidermal growth factor receptor (EGFR) in ISCs. This promotes their growth and division and, thereby, epithelial regeneration. Here we demonstrate that the HMG-box transcriptional repressor, Capicua (Cic), mediates these functions of EGFR signaling. Depleting Cic in ISCs activated them for division, whereas overexpressed Cic inhibited ISC proliferation and midgut regeneration. Epistasis tests showed that Cic acted as an essential downstream effector of EGFR/Ras signaling, and immunofluorescence showed that Cic's nuclear localization was regulated by EGFR signaling. ISC-specific mRNA expression profiling and DNA binding mapping using DamID indicated that Cic represses cell proliferation via direct targets including string (Cdc25), Cyclin E, and the ETS domain transcription factors Ets21C and Pointed (pnt). pnt was required for ISC over-proliferation following Cic depletion, and ectopic pnt restored ISC proliferation even in the presence of overexpressed dominant-active Cic. These studies identify Cic, Pnt, and Ets21C as critical downstream effectors of EGFR signaling in Drosophila ISCs.
Assuntos
Proteínas de Ligação a DNA/genética , Proteínas de Drosophila/genética , Receptores ErbB/genética , Proteínas HMGB/genética , Proteínas do Tecido Nervoso/genética , Proteínas Proto-Oncogênicas c-ets/genética , Proteínas Proto-Oncogênicas/genética , Receptores de Peptídeos de Invertebrados/genética , Proteínas Repressoras/genética , Fatores de Transcrição/genética , Animais , Proliferação de Células/genética , Proteínas de Ligação a DNA/biossíntese , Drosophila/genética , Proteínas de Drosophila/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Proteínas HMGB/biossíntese , Intestinos/citologia , Intestinos/crescimento & desenvolvimento , Proteínas do Tecido Nervoso/biossíntese , Proteínas Proto-Oncogênicas/biossíntese , RNA Mensageiro/biossíntese , Proteínas Repressoras/biossíntese , Transdução de Sinais/genética , Células-Tronco/citologia , Fatores de Transcrição/biossínteseRESUMO
Receptor Tyrosine Kinase (RTK) signaling pathways induce multiple biological responses, often by regulating the expression of downstream genes. The HMG-box protein Capicua (Cic) is a transcriptional repressor that is downregulated in response to RTK signaling, thereby enabling RTK-dependent induction of Cic targets. In both Drosophila and mammals, Cic is expressed as two isoforms, long (Cic-L) and short (Cic-S), whose functional significance and mechanism of action are not well understood. Here we show that Drosophila Cic relies on the Groucho (Gro) corepressor during its function in the early embryo, but not during other stages of development. This Gro-dependent mechanism requires a short peptide motif, unique to Cic-S and designated N2, which is distinct from other previously defined Gro-interacting motifs and functions as an autonomous, transferable repressor element. Unexpectedly, our data indicate that the N2 motif is an evolutionary innovation that originated within dipteran insects, as the Cic-S isoform evolved from an ancestral Cic-L-type form. Accordingly, the Cic-L isoform lacking the N2 motif is completely inactive in early Drosophila embryos, indicating that the N2 motif endowed Cic-S with a novel Gro-dependent activity that is obligatory at this stage. We suggest that Cic-S and Gro coregulatory functions have facilitated the evolution of the complex transcriptional network regulated by Torso RTK signaling in modern fly embryos. Notably, our results also imply that mammalian Cic proteins are unlikely to act via Gro and that their Cic-S isoform must have evolved independently of fly Cic-S. Thus, Cic proteins employ distinct repressor mechanisms that are associated with discrete structural changes in the evolutionary history of this protein family.
Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Padronização Corporal/genética , Proteínas de Drosophila/genética , Proteínas HMGB/genética , Receptores Proteína Tirosina Quinases/genética , Proteínas Repressoras/genética , Animais , Drosophila/embriologia , Drosophila/genética , Proteínas de Drosophila/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas HMGB/metabolismo , Isoformas de Proteínas/genética , Proteínas Repressoras/metabolismo , Transdução de Sinais/genéticaRESUMO
INTRODUCTION: Cardiovascular disease is the main cause of mortality worldwide. In women, its incidence increases at the sixth decade of life, coinciding with postmenopause. Whether this effect is due to menopause-related hormonal changes is not known. OBJECTIVE: To evaluate the differences in cardiovascular risk in pre- and postmenopausal women by means of the Globorisk risk scale, the triglyceride/high-density lipoproteins cholesterol (Tg/HDL-C) ratio and metabolic syndrome (MS) criteria. METHOD: Cross-sectional study that included 408 women from 40 to 60 years of age; anthropometric measurements and biochemical determinations were performed. The participants were classified as premenopausal and postmenopausal. Cardiovascular risk was assessed using the MS criteria, the Globorisk risk calculator and the Tg/HDL-C ratio. RESULTS: Postmenopausal women showed a significant increase in waist circumference, total cholesterol and triglycerides in comparison with premenopausal women. Significant associations were found between hormonal state and Globorisk measured cardiovascular risk (OR = 2.50; 95 % CI = 1.67-3.74) and the Tg/HDL-C ratio (OR = 1.66; 95 % CI = 1.09-2.52). CONCLUSION: Cardiovascular risk factors have a higher prevalence in postmenopause. The Globorisk scale and Tg/HDL-C ratio identify cardiovascular risk in postmenopausal women.
INTRODUCCIÓN: La enfermedad cardiovascular es la principal causa de mortalidad en el mundo. En la mujer se incrementa en la sexta década de la vida, coincidiendo con la posmenopausia. Se desconoce si este efecto se debe a cambios hormonales relacionados con la menopausia. OBJETIVO: Evaluar diferencias del riesgo cardiovascular en mujeres pre y posmenopáusicas mediante la escala de riesgo Globorisk, el índice triglicéridos/c-HDL (Tg/c-HDL) y los criterios de síndrome metabólico (SM). MÉTODOS: Estudio transversal que incluyó a 408 mujeres de 40 a 60 años; se realizaron mediciones antropométricas y bioquímicas. Las participantes se clasificaron en premenopáusicas y posmenopáusicas. El riesgo cardiovascular se evaluó utilizando los criterios de SM, calculadora de riesgo Globorisk y el índice Tg/c-HDL. RESULTADOS: Las mujeres en etapa posmenopáusica presentaron incremento significativo en la circunferencia de cintura, de colesterol total y triglicéridos, en comparación con las mujeres premenopáusicas. Se encontraron asociaciones significativas del estado hormonal con el riesgo cardiovascular evaluado por Globorisk (RM = 2.50, IC 95 % = 1.67-3.74) y con el índice Tg/c-HDL (RM = 1.66, IC 95 % = 1.09-2.52). CONCLUSIÓN: Los factores de riesgo cardiovascular tienen mayor prevalencia en la posmenopausia. La escala Globorisk y el índice Tg/c-HDL identifican el riesgo cardiovascular en la mujer posmenopáusica.
Assuntos
Doenças Cardiovasculares/epidemiologia , Síndrome Metabólica/epidemiologia , Pós-Menopausa , Pré-Menopausa , Adulto , Doenças Cardiovasculares/etiologia , Colesterol/sangue , HDL-Colesterol/sangue , Estudos Transversais , Feminino , Humanos , México/epidemiologia , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Triglicerídeos/sangue , Circunferência da Cintura/fisiologiaRESUMO
Objective: To establish the frequency and results in patients carriers with multiple aneurysms (MA) treated by microsurgery and/or neurological endovascular therapy (NET) in the Hospital of Specialties on The National Medical Center La Raza. Method: It is an ambispective, descriptive and longitudinal study that includes patients carriers of MA treated in the National Medical Center La Raza from March the 1st of 2009 to April the 30th of 2014. Results: 62 patients carriers of 151 aneurysms were treated. According to the type of treatment, 30 patients (49%) were included in the surgical group (GQ), 25 (40%) in the endovascular group (GE) and 7 (11%) in the combinated group (GC). The number of aneurysms was distributed this way: 69 (46%) in the GQ, 61 (40%) in the GE and 21 (14%) in the GC. At GQ, it was not possible to exclude all their aneurysms on 21% of the patients, while it was feasible in only 27%. In all GE patients (40%) the exclusion of all aneurysms was achieved. The GC, meaning surgical cases that were not completed by NET, formed 11% of the cases. At GQ there was a rate of 6% of complications, meanwhile at GE it was 0.5%.
Assuntos
Aneurisma Intracraniano/terapia , Aneurisma Roto/diagnóstico por imagem , Aneurisma Roto/epidemiologia , Aneurisma Roto/terapia , Embolização Terapêutica/métodos , Embolização Terapêutica/mortalidade , Embolização Terapêutica/estatística & dados numéricos , Procedimentos Endovasculares/métodos , Procedimentos Endovasculares/mortalidade , Procedimentos Endovasculares/estatística & dados numéricos , Feminino , Hospitais Especializados , Humanos , Aneurisma Intracraniano/diagnóstico por imagem , Aneurisma Intracraniano/epidemiologia , Estudos Longitudinais , Masculino , Microcirurgia/mortalidade , Microcirurgia/estatística & dados numéricos , Estudos Prospectivos , Estudos Retrospectivos , Distribuição por Sexo , Stents , Resultado do TratamentoRESUMO
ERK controls gene expression in development, but mechanisms that link ERK activation to changes in transcription are not well understood. We used high-resolution analysis of signaling dynamics to study transcriptional interpretation of ERK signaling during Drosophila embryogenesis, at a stage when ERK induces transcription of intermediate neuroblasts defective (ind), a gene essential for patterning of the nerve cord. ERK induces ind by antagonizing its repression by Capicua (Cic), a transcription factor that acts as a sensor of receptor tyrosine kinases in animal development and human diseases. A recent study established that active ERK reduces the nuclear levels of Cic, but it remained unclear whether this is required for the induction of Cic target genes. We provide evidence that Cic binding sites within the regulatory DNA of ind control the spatial extent and the timing of ind expression. At the same time, we demonstrate that ERK induces ind before Cic levels in the nucleus are reduced. Based on this, we propose that ERK-dependent relief of gene repression by Cic is a two-step process, in which fast reduction of repressor activity is followed by slower changes in nuclear localization and overall protein levels. This may be a common feature of systems in which ERK induces genes by relief of transcriptional repression.
Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Proteínas HMGB/metabolismo , Proteínas de Homeodomínio/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas Repressoras/metabolismo , Animais , Animais Geneticamente Modificados , Padronização Corporal/genética , Padronização Corporal/fisiologia , Núcleo Celular/genética , Núcleo Celular/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas HMGB/genética , Proteínas de Homeodomínio/genética , Humanos , Sistema de Sinalização das MAP Quinases/genética , Microfluídica , Fosforilação/fisiologia , Proteínas Repressoras/genéticaRESUMO
Dorsoventral (DV) axis formation in Drosophila begins with selective activation of EGFR, a receptor tyrosine kinase (RTK), in dorsal-anterior (DA) ovarian follicle cells. A critical event regulated by EGFR signaling is the repression of the sulfotransferase-encoding gene pipe in dorsal follicle cells, but how this occurs remains unclear. Here we show that Mirror (Mirr), a homeodomain transcription factor induced by EGFR signaling in DA follicle cells, directly represses pipe expression by binding to a conserved element in the pipe regulatory region. In addition, we find that the HMG-box protein Capicua (Cic) supports pipe expression in ventral follicle cells by repressing Mirr in this region. Interestingly, this role of Cic resembles its function in regulating anteroposterior (AP) body patterning, where Cic supports gap gene expression in central regions of the embryo by repressing Tailless, a repressor induced by RTK signaling at the embryonic poles. Thus, related RTK-Cic repressor circuits regulate the early stages of Drosophila DV and AP body axis formation.
Assuntos
Padronização Corporal/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriologia , Receptores ErbB/metabolismo , Proteínas do Olho/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas HMGB/metabolismo , Proteínas de Homeodomínio/metabolismo , Receptores de Peptídeos de Invertebrados/metabolismo , Proteínas Repressoras/metabolismo , Sulfotransferases/genética , Sulfotransferases/metabolismo , Fatores de Transcrição/metabolismo , Animais , Sequência Conservada , Proteínas de Drosophila/biossíntese , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Embrião não Mamífero/metabolismo , Feminino , Folículo Ovariano/citologia , Folículo Ovariano/embriologia , Folículo Ovariano/metabolismo , Sequências Reguladoras de Ácido Nucleico , Transdução de Sinais , Sulfotransferases/biossínteseRESUMO
The dorsoventral (DV) axis of the Drosophila embryo is patterned by a nuclear gradient of the Rel family transcription factor, Dorsal (Dl), that activates or represses numerous target genes in a region-specific manner. Here, we demonstrate that signaling by receptor tyrosine kinases (RTK) reduces nuclear levels and transcriptional activity of Dl, both at the poles and in the mid-body of the embryo. These effects depend on wntD, which encodes a Dl antagonist belonging to the Wingless/Wnt family of secreted factors. Specifically, we show that, via relief of Groucho- and Capicua-mediated repression, the Torso and EGFR RTK pathways induce expression of WntD, which in turn limits Dl nuclear localization at the poles and along the DV axis. Furthermore, this RTK-dependent control of Dl is important for restricting expression of its targets in both contexts. Thus, our results reveal a new mechanism of crosstalk, whereby RTK signals modulate the spatial distribution and activity of a developmental morphogen in vivo.
Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriologia , Drosophila melanogaster/metabolismo , Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , Receptores Proteína Tirosina Quinases/metabolismo , Fatores de Transcrição/metabolismo , Animais , Animais Geneticamente Modificados , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Padronização Corporal/genética , Padronização Corporal/fisiologia , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Receptores ErbB/genética , Receptores ErbB/metabolismo , Retroalimentação Fisiológica , Regulação da Expressão Gênica no Desenvolvimento , Genes de Insetos , Proteínas HMGB/genética , Proteínas HMGB/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Modelos Biológicos , Mutação , Proteínas Nucleares/genética , Fosfoproteínas/genética , Receptores Proteína Tirosina Quinases/genética , Receptores de Peptídeos de Invertebrados/genética , Receptores de Peptídeos de Invertebrados/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Transdução de Sinais , Fatores de Transcrição/genéticaRESUMO
Receptor tyrosine kinase (RTK) signaling pathways control multiple cellular decisions in metazoans, often by regulating the expression of downstream genes. In Drosophila melanogaster and other systems, E-twenty-six (ETS) transcription factors are considered to be the predominant nuclear effectors of RTK pathways. Here, we highlight recent progress in identifying the HMG-box protein Capicua (CIC) as a key sensor of RTK signaling in both Drosophila and mammals. Several studies have shown that CIC functions as a repressor of RTK-responsive genes, keeping them silent in the absence of signaling. Following the activation of RTK signaling, CIC repression is relieved, and this allows the expression of the targeted gene in response to local or ubiquitous activators. This regulatory switch is essential for several RTK responses in Drosophila, from the determination of cell fate to cell proliferation. Furthermore, increasing evidence supports the notion that this mechanism is conserved in mammals, where CIC has been implicated in cancer and neurodegeneration. In addition to summarizing our current knowledge on CIC, we also discuss the implications of these findings for our understanding of RTK signaling specificity in different biological processes.
Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Proteínas HMGB/metabolismo , Receptores Proteína Tirosina Quinases/fisiologia , Proteínas Repressoras/fisiologia , Transdução de Sinais/fisiologia , Animais , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Regulação Neoplásica da Expressão Gênica , Proteínas HMGB/genética , Humanos , Neoplasias/embriologia , Neoplasias/enzimologia , Neoplasias/metabolismo , Receptores Proteína Tirosina Quinases/genética , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismoRESUMO
RTK/Ras/MAPK signaling pathways play key functions in metazoan development, but how they control expression of downstream genes is not well understood. In Drosophila, it is generally assumed that most transcriptional responses to RTK signal activation depend on binding of Ets-family proteins to specific cis-acting sites in target enhancers. Here, we show that several Drosophila RTK pathways control expression of downstream genes through common octameric elements that are binding sites for the HMG-box factor Capicua, a transcriptional repressor that is downregulated by RTK signaling in different contexts. We show that Torso RTK-dependent regulation of terminal gap gene expression in the early embryo critically depends on Capicua octameric sites, and that binding of Capicua to these sites is essential for recruitment of the Groucho co-repressor to the huckebein enhancer in vivo. We then show that subsequent activation of the EGFR RTK pathway in the neuroectodermal region of the embryo controls dorsal-ventral gene expression by downregulating the Capicua protein, and that this control also depends on Capicua octameric motifs. Thus, a similar mechanism of RTK regulation operates during subdivision of the anterior-posterior and dorsal-ventral embryonic axes. We also find that identical DNA octamers mediate Capicua-dependent regulation of another EGFR target in the developing wing. Remarkably, a simple combination of activator-binding sites and Capicua motifs is sufficient to establish complex patterns of gene expression in response to both Torso and EGFR activation in different tissues. We conclude that Capicua octamers are general response elements for RTK signaling in Drosophila.
Assuntos
Proteínas de Ligação a DNA , Proteínas de Drosophila/genética , Proteínas HMGB/genética , Sistema de Sinalização das MAP Quinases , Receptores Proteína Tirosina Quinases/metabolismo , Proteínas Repressoras/genética , Elementos de Resposta , Animais , Sítios de Ligação , Padronização Corporal , Drosophila , Receptores ErbB/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Multimerização Proteica , Asas de Animais/crescimento & desenvolvimentoRESUMO
Pollution in aquatic ecosystems has been increasing drastically worldwide changing their water quality. Therefore, species must be adapted to these new scenarios. In Aguascalientes City, four representative urban reservoirs contain lead in the water column and extremely high concentrations of sediments. Therefore, an analysis was conducted to evaluate the resilience of zooplankton species to lead exposure in each reservoir using dormant and organisms. Results demonstrated a decrease range from 57.5 to 22.5% in overall diapausing egg hatching rate, while survivorship rate also decreased from 98 to 54% when organisms were exposed to the water of the four reservoirs and increasing lead concentrations. When Pb exposure increased, results showed a global negative effect on both hatching rate (decreasing from 58 to 30% at 0.09 mg L-1) and survivorship levels (decreasing from 100% to 0.07% at 0.09 mg L-1). We provide Species Sensitivity Distribution for both water reservoir dilutions and lead concentration to analyze diapausing eggs hatching and survivorship of offspring in the presence of same polluted conditions or lead of the autochthonous species found in reservoirs. Furthermore, specific analysis with two populations of the cladoceran Moina macrocopa showed clear dissimilar hatching patterns that suggested a different adaptive mechanism. Niagara population shows a hatching rate of approximately 25% in the first two days of reservoir water exposure, while UAA population drastically increased hatching rate to 75% on exposure at day seven. We provide the first record of bioaccumulation in ephippia of M. macrocopa.
Assuntos
Cladocera , Resiliência Psicológica , Rotíferos , Poluentes Químicos da Água , Animais , Chumbo/toxicidade , Ecossistema , México , Poluentes Químicos da Água/toxicidade , Eutrofização , ZooplânctonRESUMO
Diapausing embryos encased within cladoceran ephippia result from sexual reproduction and increase genetic diversity. They are also important means by which species bypass harsh environmental conditions and disperse in space and time. Once released, ephippia usually sink to the benthos and remain there until hatching. Using the Sars' method (incubating sediments to identify cladoceran hatchlings), ephippial egg bank biodiversity can be evaluated. Yet, even when samples are incubated under a variety of conditions, it is not possible to warrant that all have hatched. Few keys are available that facilitate the identification of cladocerans by using only ephippial morphology. Our goal was to analyze some cladoceran ephippia from Mexico, to develop a means to identify them using easily recognizable characteristics. Ephippia of 23 cladoceran species from waters in Aguascalientes (México) in 11 genera (Alona, Biapertura, Ceriodaphnia, Chydorus, Daphnia, Dunhevedia, Ilyocryptus, Macrothrix, Moina, Pleuroxus, and Simocephalus) were analyzed. In our analysis six morphological features were selected that permitted the identification of ephippia to species(-group) level. The results demonstrate that with a proper catalog of features, some ephippia can be identified.
RESUMO
The study of sexual reproductive behavior supported by ultrastructural evidence is important in rotifers to describe differences among potential cryptic species. In this research, the morphology of the rotifer Brachionus bidentatus is described at the ultrastructural level, using electronic microscopy, together with a brief description and discussion of its sexual reproductive behavior. The characteristics of the (a) male, (b) the female, (c) the sexual egg or cyst, (d) the partenogenic egg, (e) the no-fecundated sexual egg (male egg), and (f) the trophi, were described. Another part of this research is dedicated to the ultrastructure of the sex cells of the male rotifer B. bidentatus. Samples were obtained from La Punta pond in Cosio, Aguascalientes, Mexico (22 degrees 08' N - 102 degrees 24' W), and a culture was maintained in the laboratory. Fifty organisms, from different stages of the rotifer Brachionus bidentatus, were fixed in Formol at 4% and then prepared; besides, for the trophi, 25 female rotifer Brachionus bidentatus were prepared for observation in a JEOL 5900 LV scanning electronic microscope. In addition, for the observation of male sex cells, 500 males of Brachionus bidentatus were isolated, fixed and observed in a JEOL 1010 transmission microscope. Females of B. bidentatus in laboratory cultures had a lifespan of five days (mean+one SD = 4.69 +/- 0.48; N=13), and produced 4.5 +/- 3.67 (N=6) parthenogenetic eggs during such lifespan. In the case of non-fertilized sexual eggs, they produced up to 18 eggs (mean+one SD = 13 +/- 4.93; N=7). Sexual females produced a single cyst on average (mean +/- one SD = I +/- 0; N=20). For the sexual cycle, the time of copulation between male and female ranged from 10 to 40 seconds (mean +/- one SD = 17.33 +/- 10.55, N=7). The spermatozoa are composed of a celular body and a flagellum, the size of the body is of 300 nm while the flagellum measures 1 700nm. The rods have a double membrane. Their mean length is almost 2.45 microm +/- 0.74, N=6; and their mean wide is 0.773 microm +/- 0.241, N=11. The evidence on the specific ultrastructural characteristics of the rotifer B. bidentatus is notorious, even more in the male and in the cyst cell. Regarding the ultrastructure of the spermatozoa and the rods, compared to other species they only differ in size, despite their structural resemblance. Our study of the ultraestructure of this species adds useful information that along with molecular data will help clarify the taxonomy of brachionid rotifers.
Assuntos
Óvulo/ultraestrutura , Rotíferos/ultraestrutura , Espermatozoides/diagnóstico por imagem , Animais , Feminino , Masculino , México , Microscopia Eletrônica de Transmissão , Reprodução , Rotíferos/anatomia & histologia , UltrassonografiaRESUMO
Background: Blood loss estimation in a surgery is made by anesthesiologists by means of visual technique, which is not reliable because it can change depending on the judgement of every person, or his/her work experience, which is why it is considered something subjective. Therefore, the results obtained could lead to make mistakes with the exact amount of bleeding, mismanaging unnecessary hemoderived transfusions or administering unnecessary drugs. Objective: To compare the blood volume and its visual calculation between Anesthesiology residents and anesthesiologists. Material and methods: Prolective cross-sectional study which included 85 Anesthesiology residents and anesthesiologists. Diverse scenarios of bleeding were set, divided into gauze pads, compresses and jars, and each participant was asked to answer the poll. Results: There is no significant difference in the estimate of bleeding between the estimation made by residents and anesthesiologists. Conclusions: The grade of studies or experience is not significant for the exact estimation of bleeding with visual technique.
Introducción: la estimación de sangrado en las cirugías se realiza por medio de la técnica de cálculo visual, que es llevada a cabo por parte del servicio de anestesiología. Se trata de una técnica poco confiable, que puede variar dependiendo del juicio de cada persona y de su experiencia laboral, por lo que es considerada algo subjetivo. Por lo tanto, los datos arrojados hacen que se cometan errores en la cantidad exacta del sangrado y llevan a transfusiones de paquetes globulares o a la administración probablemente innecesaria de medicamentos. Objetivo: comparar la cantidad de un volumen de sangrado determinado y su cálculo visual realizada por residentes de Anestesiología y anestesiólogos. Material y métodos: estudio transversal prolectivo en el cual se incluyeron 85 médicos residentes de Anestesiología y anestesiólogos. Se dispusieron diversos escenarios con sangrado, los cuales se dividieron en gasas, compresas y frascos, y se le pidió a cada médico que contestara la encuesta correspondiente según su criterio. Resultados: de acuerdo con los resultados, no hay diferencia significativa en la estimación realizada del sangrado entre residentes y anestesiólogos para las muestras estudiadas. Conclusiones: el grado de estudios o experiencia no es significativo para la exacta estimación de sangrado con técnica visual.