RESUMEN
Long-read sequencing (LRS) technologies have provided extremely powerful tools to explore genomes. While in the early years these methods suffered technical limitations, they have recently made significant progress in terms of read length, throughput, and accuracy and bioinformatics tools have strongly improved. Here, we aim to review the current status of LRS technologies, the development of novel methods, and the impact on genomics research. We will explore the most impactful recent findings made possible by these technologies focusing on high-resolution sequencing of genomes and transcriptomes and the direct detection of DNA and RNA modifications. We will also discuss how LRS methods promise a more comprehensive understanding of human genetic variation, transcriptomics, and epigenetics for the coming years.
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Genómica , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Genómica/métodos , Análisis de Secuencia de ADN/métodos , Biología Computacional , Perfilación de la Expresión Génica/métodosRESUMEN
Access to accurate viral genomes is important to downstream data analysis. Third-generation sequencing (TGS) has recently become a popular platform for virus sequencing because of its long read length. However, its per-base error rate, which is higher than next-generation sequencing, can lead to genomes with errors. Polishing tools are thus needed to correct errors either before or after sequence assembly. Despite promising results of available polishing tools, there is still room to improve the error correction performance to perform more accurate genome assembly. The errors, particularly those in coding regions, can hamper analysis such as linage identification and variant monitoring. In this work, we developed a novel pipeline, HMMPolish, for correcting (polishing) errors in protein-coding regions of known RNA viruses. This tool can be applied to either raw TGS reads or the assembled sequences of the target virus. By utilizing profile Hidden Markov Models of protein families/domains in known viruses, HMMPolish can correct errors that are ignored by available polishers. We extensively validated HMMPolish on 34 datasets that covered four clinically important viruses, including HIV-1, influenza-A, norovirus, and severe acute respiratory syndrome coronavirus 2. These datasets contain reads with different properties, such as sequencing depth and platforms (PacBio or Nanopore). The benchmark results against popular/representative polishers show that HMMPolish competes favorably on error correction in coding regions of known RNA viruses.
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COVID-19 , Virus ARN , Virus , Humanos , Análisis de Secuencia de ADN/métodos , Genoma , Secuenciación de Nucleótidos de Alto Rendimiento/métodosRESUMEN
PIWI-interacting RNAs (piRNAs), which are germ cell-specific small RNAs, are essential for spermatogenesis. In fetal mouse germ cells, piRNAs are synthesized from sense and antisense RNAs of transposable element sequences for retrotransposon silencing. In a previous study, we reported that transgenic mice expressing antisense-Dnmt3L under the control of the Miwi2 promoter (Tg-Miwi2P-asDnmt3L) exhibited piRNA-mediated DNMT3L down-regulation. In this study, two transgene integration loci (B3 and E1) were identified on chromosome 18 of the Tg-Miwi2P-asDnmt3L mice; these loci were weak piRNA clusters. Crossbreeding was performed to obtain mice with the transgene cassette inserted into a single locus. DNMT3L was silenced and spermatogenesis was severely impaired in mice with the transgene cassette inserted at the B3 locus (Tg-B mice). In contrast, spermatogenesis in mice bearing the transgene at the E1 locus (Tg-E mice) was normal. The number of piRNAs for Dnmt3L in Tg-B mice was eightfold higher than that in Tg-E mice. Therefore, both gene silencing and impaired spermatogenesis depended on the transgene copy number rather than on the insertion loci. Additionally, the endogenous Dnmt3L promoter was not methylated in Tg mice, suggesting that Dnmt3L silencing was caused by post-transcriptional gene silencing. Based on these data, we discuss a piRNA-dependent gene silencing mechanism against novel gene insertions.
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Variaciones en el Número de Copia de ADN , Silenciador del Gen , Animales , Proteínas Argonautas/genética , ADN (Citosina-5-)-Metiltransferasas/genética , Masculino , Ratones , ARN Interferente Pequeño/genética , Espermatogénesis/genética , Factores de Transcripción/genética , TransgenesRESUMEN
BACKGROUND: Thalassemias represent some of the most common monogenic diseases worldwide and are caused by variations in human hemoglobin genes which disrupt the balance of synthesis between the alpha and beta globin chains. Thalassemia gene detection technology is the gold standard to achieve accurate detection of thalassemia, but in clinical practice, most of the tests are only for common genotypes, which can easily lead to missing or misdiagnosis of rare thalassemia genotypes. CASE PRESENTATION: We present the case of an 18-year-old Chinese female with abnormal values of routine hematological indices who was admitted for genetic screening for thalassemia. Genomic DNA was extracted and used for the genetic assays. Gap polymerase chain reaction and agarose gel electrophoresis were performed to detect HBA gene deletions, while PCR-reverse dot blot hybridization was used to detect point mutations in the HBA and HBB genes. Next-generation sequencing and third-generation sequencing (TGS) were used to identify known and potentially novel genotypes of thalassemia. We identified a novel complex variant αHb WestmeadαHb Westmeadαanti3.7/-α3.7 in a patient with rare alpha-thalassemia. CONCLUSIONS: Our study identified a novel complex variant that expands the thalassemia gene variants spectrum. Meanwhile, the study suggests that TGS could effectively improve the specificity of thalassemia gene detection, and has promising potential for the discovery of novel thalassemia genotypes, which could also improve the accuracy of genetic counseling. Couples who are thalassemia carriers have the opportunity to reduce their risk of having a child with thalassemia.
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Talasemia alfa , Humanos , Talasemia alfa/genética , Talasemia alfa/diagnóstico , Femenino , Adolescente , Secuenciación de Nucleótidos de Alto Rendimiento , Genotipo , Pruebas Genéticas/métodos , Mutación Puntual , Hemoglobinas Anormales/genéticaRESUMEN
In our studies, we combined two powdered materials, i.e., ferroelectric triglycine sulfate (TGS) and ferrimagnetic magnetite Fe3O4, to obtain a magnetoelectric composite. The ferroelectric (E) part, i.e., TGS, was a hybrid organic-inorganic crystal, which we obtained as a pure single crystal from an aqueous solution using a static water evaporation method. The magnetic (M) part of the composite was commercially available magnetite. The samples used for the dielectric and magnetoelectric measurements were cold-pressed and made in the form of a circular tablet. The measuring electrodes were made of silver-based conductive paste and were attached to the sample. We measured the temperature dependencies of selected electrical parameters (e.g., dielectric permittivity, electrical capacity, and loss angle tangent). We used the dynamic lock-in method to check whether magnetoelectric coupling existed between the E and M phases. In this paper, we present the dielectric properties of pure monocrystalline TGS as a reference sample and compare the results for TGS powder, TGS + carbon powder, and TGS + Fe3O4 powder. The magnetoelectric coupling presumably appeared for the composite TGS + 10 wt. % Fe3O4, as evidenced by the shift in the phase transition temperature in the TGS. Moreover, the theoretical interpretation of the effect is proposed.
RESUMEN
Here, we report the characterization of a plant RNA methyltransferase, orthologous to yeast trimethylguanosine synthase1 (Tgs1p) and whose downregulation was associated with apomixis in Paspalum grasses. Using phylogenetic analyses and yeast complementation, we determined that land plant genomes all encode a conserved, specific TGS1 protein. Next, we studied the role of TGS1 in female reproduction using reporter lines and loss-of-function mutants in Arabidopsis thaliana. pAtTGS1:AtTGS1 reporters showed a dynamic expression pattern. They were highly active in the placenta and ovule primordia at emergence but, subsequently, showed weak signals in the nucellus. Although expressed throughout gametophyte development, activity became restricted to the female gamete and was also detected after fertilization during embryogenesis. TGS1 depletion altered the specification of the precursor cells that give rise to the female gametophytic generation and to the sporophyte, resulting in the formation of a functional aposporous-like lineage. Our results indicate that TGS1 participates in the mechanisms restricting cell fate acquisition to a single cell at critical transitions throughout the female reproductive lineage and, thus, expand our current knowledge of the mechanisms governing female reproductive fate in plants.
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Arabidopsis , Arabidopsis/metabolismo , Saccharomyces cerevisiae , Filogenia , Mutación/genética , Óvulo Vegetal/metabolismo , Células Germinativas , Regulación de la Expresión Génica de las PlantasRESUMEN
We investigated whether 20 candidate single nucleotide polymorphisms (SNPs) were associated with in vivo exercise-induced muscle damage (EIMD), and with an in vitro skeletal muscle stem cell wound healing assay. Sixty-five young, untrained Caucasian adults performed 120 maximal eccentric knee-extensions on an isokinetic dynamometer to induce EIMD. Maximal voluntary isometric/isokinetic knee-extensor torque, knee joint range of motion (ROM), muscle soreness, serum creatine kinase activity and interleukin-6 concentration were assessed before, directly after and 48 h after EIMD. Muscle stem cells were cultured from vastus lateralis biopsies from a separate cohort (n = 12), and markers of repair were measured in vitro. Participants were genotyped for all 20 SNPs using real-time PCR. Seven SNPs were associated with the response to EIMD, and these were used to calculate a total genotype score, which enabled participants to be segregated into three polygenic groups: 'preferential' (more 'protective' alleles), 'moderate', and 'non-preferential'. The non-preferential group was consistently weaker than the preferential group (1.93 ± 0.81 vs. 2.73 ± 0.59 N â m/kg; P = 9.51 × 10-4 ) and demonstrated more muscle soreness (p = 0.011) and a larger decrease in knee joint ROM (p = 0.006) following EIMD. Two TTN-AS1 SNPs in linkage disequilibrium were associated with in vivo EIMD (rs3731749, p ≤ 0.005) and accelerated muscle stem cell migration into the artificial wound in vitro (rs1001238, p ≤ 0.006). Thus, we have identified a polygenic profile, linked with both muscle weakness and poorer recovery following EIMD. Moreover, we provide evidence for a novel TTN gene-cell-skeletal muscle mechanism that may help explain some of the interindividual variability in the response to EIMD.
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Ejercicio Físico , Músculo Esquelético/fisiología , Mialgia , Adulto , Ejercicio Físico/fisiología , Humanos , Músculo Esquelético/patología , Mialgia/genética , Mialgia/patología , Polimorfismo de Nucleótido Simple , Músculo Cuádriceps/citología , Músculo Cuádriceps/fisiología , Células Madre/citología , TorqueRESUMEN
In fission yeast, siRNA generated by RNA interference (RNAi) factors plays critical roles in establishment and maintenance of heterochromatin. To achieve efficient siRNA synthesis, RNAi factors assemble on heterochromatin via association with Swi6, a homologue of heterochromatin protein 1 (HP1), and heterochromatic noncoding RNA (hncRNA) retained on chromatin. In addition, spliceosomes formed on hncRNA introns recruit RNAi factors to hncRNA and heterochromatin. Small nuclear RNAs, components of the spliceosome, have a trimethylguanosine (TMG) cap that is generated by Tgs1-dependent hypermethylation of the normal m7G cap; this cap is required for efficient splicing of some mRNAs in budding yeast and Drosophila. In this study, we found that loss of Tgs1 in fission yeast destabilizes centromeric heterochromatin. Tgs1 was required for Swi6-independent siRNA synthesis, as well as for the establishment of centromeric heterochromatin. Loss of Tgs1 affected the splicing efficiency of hncRNA introns in the absence of Swi6. Furthermore, some hncRNAs have a TMG cap, and we found that loss of Tgs1 diminished the chromatin binding of these hncRNAs. Together, these results suggest that the Tgs1-dependent TMG cap plays critical roles in establishment of heterochromatin by ensuring spliceosome-dependent recruitment of RNAi factors and regulating the binding between chromatin and hncRNA.
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Proteínas Cromosómicas no Histona/metabolismo , Heterocromatina/metabolismo , ARN Interferente Pequeño/metabolismo , Proteínas de Schizosaccharomyces pombe/metabolismo , Schizosaccharomyces/metabolismo , ARNt Metiltransferasas/metabolismo , Centrómero/metabolismo , Silenciador del Gen , Intrones/genética , Modelos Biológicos , Dominios Proteicos , Empalme del ARN/genética , ARN sin Sentido/metabolismo , ARN de Hongos/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas de Schizosaccharomyces pombe/química , ARNt Metiltransferasas/químicaRESUMEN
BACKGROUND AND AIM: Serum lipids, glucose and uric acid are well-known risk factors for metabolic syndrome and cardiovascular diseases; however, how serum uric acid levels are associated with fasting glucose and lipid levels remains to be evaluated. METHODS AND RESULTS: A cross-sectional study was performed in 104,328 males and 74,916 females. Quantile regression analyses were adopted to optimally fit the associations between levels of uric acid, lipids and glucose. Fasting high-density lipoprotein cholesterol (HDL-C) levels were negatively associated with serum uric acid levels; the associations remained stable in males but strengthened in females with increasing uric acid concentrations. Non-HDL-C and triglyceride (TG) levels were positively associated with serum uric acid levels; the associations also strengthened across deciles of uric acid levels from low to high. Fasting glucose levels were positively associated with uric acid levels in both sexes except in males in the 1st and 2nd deciles of uric acid concentrations; the association coefficients for females were higher than coefficients in males in each decile of uric acid levels. All associations had distinguishable patterns by sex except non-HDL-C, which was associated with uric acid levels with relatively similar trends between sexes. Adjustment for known confounding factors only slightly altered the above associations. CONCLUSIONS: Fasting serum uric acid levels are associated with fasting levels of HDL-C, non-HDL-C, TG and glucose; the associations strengthened with the deciles of uric acid increase and displayed non-negligible sex differences.
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Ayuno , Ácido Úrico , Humanos , Femenino , Masculino , Triglicéridos , Glucosa , Estudios Transversales , Índice de Masa Corporal , HDL-Colesterol , Colesterol , Lipoproteínas , Glucemia/metabolismoRESUMEN
Novel pyrrolo [2,3-b] pyrrole derivatives were synthesized and their hypolipidemic activity was assessed in hyperlipidemic rats. The chemical structures of the new derivatives were confirmed through spectral analysis. Compounds 5 and 6 were revealed to be the most effective hypolipidemic agents, with considerable hypocholesterolemic and hypotriglyceridemic effects. They appear to be promising candidates for creating new powerful derivatives with anti-atherosclerotic and hypolipidemic properties. As for antimicrobial activity, some of the tested compounds showed moderate activity against Pseudomonas aeruginosa: compound 2 revealed an MIC value of 50 µg/mL, compared to 25 µg/mL for ciprofloxacin. Compound 3 showed good antimicrobial activity against Staphylococcus aureus, comparable to ciprofloxacin, and roughly half the activity of ampicillin, according to MIC values. Compound 2 has an MIC approximately 25% of that of clotrimazole against Candida albicans. Compound 2 also showed the highest antioxidant activity with 59% inhibition of radical scavenging activity. Additionally, the cytotoxic activity of these new derivatives 1-7 was investigated and most of them showed good anticancer activity against the three tested cell lines.
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Antiinfecciosos , Pirroles , Animales , Antibacterianos/química , Antiinfecciosos/farmacología , Ciprofloxacina , Pruebas de Sensibilidad Microbiana , Microondas , Simulación del Acoplamiento Molecular , Estructura Molecular , Pirroles/farmacología , Ratas , Relación Estructura-ActividadRESUMEN
Bacteria respond to nutritional stresses by changing the cellular concentration of the alarmone (p)ppGpp. This control mechanism, called the stringent response, depends on two enzymes, the (p)ppGpp synthetase RelA and the bifunctional (p)ppGpp synthetase/hydrolase SpoT in Escherichia coli and related bacteria. Because SpoT is the only enzyme responsible for (p)ppGpp hydrolysis in these bacteria, SpoT activity needs to be tightly regulated to prevent the uncontrolled accumulation of (p)ppGpp, which is lethal. To date, however, no such regulation of SpoT (p)ppGpp hydrolase activity has been documented in E. coli In this study, we show that Rsd directly interacts with SpoT and stimulates its (p)ppGpp hydrolase activity. Dephosphorylated HPr, but not phosphorylated HPr, of the phosphoenolpyruvate-dependent sugar phosphotransferase system could antagonize the stimulatory effect of Rsd on SpoT (p)ppGpp hydrolase activity. Thus, we suggest that Rsd is a carbon source-dependent regulator of the stringent response in E. coli.
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Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Guanosina Pentafosfato/metabolismo , Pirofosfatasas/metabolismo , Proteínas Represoras/metabolismo , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Guanosina Pentafosfato/genética , Pirofosfatasas/genética , Proteínas Represoras/genéticaRESUMEN
PURPOSE: To investigate use of the third-generation sequencing (TGS) Oxford Nanopore system as a new approach for preimplantation genetic testing (PGT). METHODS: Embryos with known structural variations underwent multiple displacement amplification to create fragments of DNA (average ~ 5 kb) suitable for sequencing on a nanopore. RESULTS: High-depth sequencing identified the deletion interval for the relatively large HBA1/2--SEA alpha thalassemia deletion. In addition, STRs were able to be identified in the primary sequence data for potential use in conventional PGT-M linkage confirmation. Sequencing of amplified embryo DNA carrying a translocation enabled balanced embryos to be identified and gave the precise identification of translocation breakpoints, offering the opportunity to differentiate carriers from non-carrier embryos. Low-pass sequencing gave reproducible profiles suitable for simple identification of whole-chromosome and segmental aneuploidies. CONCLUSION: TGS on the Oxford Nanopore is a possible alternative and versatile approach to PGT with potential for performing economical workups where the long read sequencing information can be used for assisting in a traditional PGT workup to design an accurate and reliable test. Additionally, application of TGS has the possibility of providing combined PGT-A/SR or in selected stand-alone PGT-M cases involving pathogenic deletions. Both of these applications offer the opportunity for simultaneous aneuploidy detection to select either balanced embryos for transfer or additional carrier identification. The low cost of the instrument offers new laboratories economical entry into onsite PGT.
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Pruebas Genéticas , Secuenciación de Nucleótidos de Alto Rendimiento/tendencias , Diagnóstico Preimplantación/tendencias , Translocación Genética/genética , Aneuploidia , Blastocisto/metabolismo , Transferencia de Embrión/métodos , Femenino , Fertilización In Vitro/tendencias , Humanos , EmbarazoRESUMEN
The PIWI-interacting RNA (piRNA) pathway is a conserved defense mechanism that protects the genetic information of animal germ cells from the deleterious effects of molecular parasites, such as transposons. Discovered nearly a decade ago, this small RNA silencing system comprises PIWI-clade Argonaute proteins and their associated RNA-binding partners, the piRNAs. In this review, we highlight recent work that has advanced our understanding of how piRNAs preserve genome integrity across generations. We discuss the mechanism of piRNA biogenesis, give an overview of common themes as well as differences in piRNA-mediated silencing between species, and end by highlighting known and emerging functions of piRNAs.
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Proteínas Argonautas/genética , Drosophila melanogaster/genética , Silenciador del Gen , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Animales , Proteínas Argonautas/metabolismo , Proteínas Cromosómicas no Histona/genética , Proteínas Cromosómicas no Histona/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Histonas/genética , Histonas/metabolismo , Humanos , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/metabolismo , ARN Mensajero/metabolismo , ARN Interferente Pequeño/metabolismo , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Transcripción GenéticaRESUMEN
In plants, RNA-directed DNA methylation (RdDM)-mediated transcriptional gene silencing (TGS) is a natural antiviral defense against geminiviruses. Several geminiviral proteins have been shown to target the enzymes related to the methyl cycle or histone modification; however, it remains largely unknown whether and by which mechanism geminiviruses directly inhibit RdDM-mediated TGS. In this study, we showed that Cotton leaf curl Multan virus (CLCuMuV) V2 directly interacts with Nicotiana benthamiana AGO4 (NbAGO4) and that the L76S mutation in V2 (V2L76S) abolishes such interaction. We further showed that V2, but not V2L76S, can suppresses RdDM and TGS. Silencing of NbAGO4 inhibits TGS, reduces the viral methylation level, and enhances CLCuMuV DNA accumulation. In contrast, the V2L76S substitution mutant attenuates CLCuMuV infection and enhances the viral methylation level. These findings reveal that CLCuMuV V2 contributes to viral infection by interaction with NbAGO4 to suppress RdDM-mediated TGS in plants.IMPORTANCE In plants, the RNA-directed DNA methylation (RdDM) pathway is a natural antiviral defense mechanism against geminiviruses. However, how geminiviruses counter RdDM-mediated defense is largely unknown. Our findings reveal that Cotton leaf curl Multan virus V2 contributes to viral infection by interaction with NbAGO4 to suppress RNA-directed DNA methylation-mediated transcriptional gene silencing in plants. Our work provides the first evidence that a geminiviral protein is able to directly target core RdDM components to counter RdDM-mediated TGS antiviral defense in plants, which extends our current understanding of viral counters to host antiviral defense.
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Geminiviridae/genética , Silenciador del Gen/fisiología , Transcripción Genética/genética , Proteínas Virales/genética , Begomovirus/genética , Metilación de ADN/genética , ADN Viral/genética , Interacciones Huésped-Patógeno/genética , Enfermedades de las Plantas/virología , Nicotiana/virologíaRESUMEN
Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), can persist in the host for decades without causing TB symptoms and can cause a latent infection, which is an intricate challenge of current TB control. The DosR regulon, which contains approximately 50 genes, is crucial in the non-replicating persistence of Mtb. tgs1 is one of the most powerfully induced genes in this regulon during Mtb non-replicating persistence. The gene encodes a triacyl glycerol synthase catalyzing synthesis of triacyl glycerol (TAG), which is proposed as an energy source during bacilli persistence. Here, western blotting showed that the Tgs1 protein was upregulated in clinical Mtb strains. To detect its physiological effects on mycobacterium, we constructed serial recombinant M. marinum including over-expressed Tgs1(Tgs1-H), reduced-expressed Tgs1(Tgs1-L), and wild type M. marinum strains as controls. Tgs1 over-expression did not influence M. marinum growth under aerobic shaking and in hypoxic cultures, while growth advantages were observed at an early stage under nutrient starvation. Transmission electron microscopy revealed more lipid droplets in Tgs1-H than the other two strains; the droplets filled the cytoplasm. Two-dimensional thin-layer chromatography revealed more phosphatidyl-myo-inositol mannosides in the Tgs1-H cell wall. To assess the virulence of recombinant M. marinum in the natural host, adult zebrafish were infected with Tgs1-H or wild type strains. Hypervirulence of Tgs1-H was characterized by markedly increased bacterial load and early death of adult zebrafish. Remarkably, zebrafish infected with Tgs1-H developed necrotizing granulomas much more rapidly and in higher amounts, which facilitated mycobacterial replication and dissemination among organs and eventual tissue destruction in zebrafish. RNA sequencing analysis showed Tgs1-H induced 13 genes differentially expressed under aerobiosis. Among them, PE_PGRS54 (MMAR_5307),one of the PE_PGRS family of antigens, was markedly up-regulated, while 110 coding genes were down-regulated in Tgs1-L.The 110 genes included 22 member genes of the DosR regulon. The collective results indicate an important role for the Tgs1 protein of M. marinumin progression of infection in the natural host. Tgs1 signaling may be involved in a previously unknown behavior of M. marinum under hypoxia/aerobiosis.
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Proteínas Bacterianas/genética , Interacciones Huésped-Patógeno , Mycobacterium marinum/genética , Mycobacterium marinum/patogenicidad , Pez Cebra/microbiología , Aerobiosis , Animales , Hipoxia , Macrófagos/microbiología , Infecciones por Mycobacterium no Tuberculosas/microbiología , Regulón , Transducción de Señal , Transcriptoma , Regulación hacia Arriba , VirulenciaRESUMEN
BACKGROUND: The present study aimed to assess the relation between nutrient patterns and changes in adult anthropometric and cardiometabolic factors. METHODS: This study was conducted on 1637 adults participating in the Tehran Lipid and Glucose Study (2005-2008), who were free of cardiovascular diseases and cancer and had completed dietary data. They were followed to the next survey (2008-2011). Dietary intakes were collected and nutrient patterns were obtained. Three year changes in anthropometric and cardiometabolic factors were measured. RESULTS: Five nutrient patterns were extracted. The first pattern was characterized by "plant protein, thiamine, niacin, and minerals including phosphorus, zinc, copper, magnesium, manganese, and selenium". Animal protein, lactose, vitamin D, riboflavine, pantothenic acid, vitamin B12, calcium, phosphorus, and zinc" were loaded in the second pattern. The third and fourth patterns were characterized by "vitamin K, fiber, calcium, iron, manganese, and potassium", and "high correlation with starch, thiamine and folate, and negative correlation with mono and poly unsaturated fatty acids and vitamin E", respectively. The fifth pattern was high in Fructose, vitamins A, C, pyridoxine, and potassium. There was no association between nutrient patterns and 3-year changes in blood pressure and fasting blood glucose; whereas, per each quartile increment of the fifth pattern adjusted for potential confounders, triglyceride change was decreased [ß = - 3.66, 95% CI (- 6.57, - 0.57); P for trend = 0.014]. CONCLUSION: Present study indicates that nutrient patterns may have an association with cardiometabolic factors, particularly a pattern rich in fructose, vitamins A, C, pyridoxine, and potassium which decreases triglyceride level.
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Enfermedades Cardiovasculares/etiología , Dieta/efectos adversos , Enfermedades Metabólicas/etiología , Nutrientes/análisis , Adulto , Antropometría , Dieta/estadística & datos numéricos , Encuestas sobre Dietas , Fibras de la Dieta/análisis , Ácidos Grasos Insaturados/análisis , Femenino , Humanos , Irán , Masculino , Persona de Mediana Edad , Minerales/análisis , Factores de Riesgo , Triglicéridos/sangre , Vitaminas/análisisRESUMEN
Cytosine DNA methylation is a conserved epigenetic silencing mechanism that defends against biotic stresses such as geminivirus infection. As a countermeasure, geminiviruses encode proteins that inhibit methylation and transcriptional gene silencing (TGS). Previous studies showed that V2 protein of Tomato yellow leaf curl virus (TYLCV) functions as a TGS suppressor. However, how V2 mediates TGS suppression remains unknown. Here we show that V2 interacts directly with a Nicotiana benthamiana histone deacetylase 6 (NbHDA6), a homolog of Arabidopsis HDA6 (AtHDA6), known to be involved in gene silencing in cooperation with methyltransferase 1 (MET1). NbHDA6 genetically complemented a late-flowering phenotype and restored histone deacetylation of an AtHDA6 mutant. Furthermore, our investigation showed that NbHDA6 displayed histone deacetylase enzymatic activity, which was not inhibited by V2. Genetic analysis revealed that silencing of NbHDA6 expression resulted in enhanced susceptibility to TYLCV infection. In addition, methylation-sensitive PCR and bisulfite sequencing analysis showed that silencing of NbHDA6 expression caused reduced DNA methylation of the viral genome in infected plants. HDA6 was previously shown to recruit and physically interact with MET1 to function in gene silencing. Using competitive pulldown and coimmunoprecipitation assays, we demonstrated that V2 did not interact but competed with NbMET1 for direct binding to NbHDA6. These findings suggest that V2 interacts with host HDA6 and interferes with the recruitment of MET1 by HDA6, resulting in decreased methylation of the viral DNA genome by TGS with a concomitant increase in host susceptibility to TYLCV infection.IMPORTANCE Plants employ repressive viral genome methylation as an epigenetic defense against geminiviruses. In turn, geminiviruses encode proteins that inhibit methylation by TGS. Previous studies showed that TYLCV V2 can efficiently suppress TGS, but the mechanism remains unknown. We showed that V2 interacted with NbHDA6 but did not inhibit its enzymatic activity. As HDA6 is known to be involved in gene silencing in cooperation with MET1, we explored the relationship between V2, NbMET1, and NbHDA6. Our investigation showed that V2 did not interact but competed with NbMET1 for direct binding to NbHDA6. To our knowledge, this is the first report that viral proteins inhibit TGS by interacting with histone deacetylase but not by blocking the methyl cycle. This work provides an additional mechanism for TGS suppression by geminiviruses.
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Begomovirus/genética , Metilación de ADN , Silenciador del Gen , Histona Desacetilasa 6/genética , Nicotiana/virología , Proteínas Virales/genética , Begomovirus/metabolismo , Genoma Viral , Histona Desacetilasa 6/metabolismo , Interacciones Huésped-Patógeno/genética , Nicotiana/enzimología , Nicotiana/genética , Proteínas Virales/metabolismoRESUMEN
To explore the approaches and diagnostic yield of genetic testing for renal disease in children, we describe the genotype and phenotype of the national cohort of children with renal disease from 13 different regions of China recruited from 2014 to 2018 by building up the multicenter registration system (Chinese Children Genetic Kidney Disease Database, CCGKDD). Genetic diagnosis was confirmed in 42.1% of our cohort of 1001 pediatric patients with clinical suspicion of a genetic renal disease. Of the 106 distinct monogenetic disorders detected, 15 accounted for 60.7% of genetic diagnoses. The diagnostic yield was 29.1% in steroid resistant nephritic syndrome (SRNS), 61.4% in cystic renal disease, 17.0% in congenital anomalies of the kidney and urinary tract (CAKUT), 62.3% in renal tubular disease/renal calcinosis, and 23.9% for chronic kidney disease (CKD) 3 to 5 stage with unknown origin. Genetic approaches of target gene sequence (TGS), singleton whole-exome sequencing (WES) and trio-WES were performed with diagnostic rates of 44.8%, 36.2%, and 42.6%, respectively. The early use of trio-WES could improve the diagnostic rate especially in renal tubular disease and calcinosis. We report the genetic spectrum of Chinese children with renal disease. Establishment of the CCGKDD will improve the genetic work on renal disease.
Asunto(s)
Exoma/genética , Predisposición Genética a la Enfermedad , Enfermedades Renales Quísticas/genética , Insuficiencia Renal Crónica/genética , Niño , Preescolar , China/epidemiología , Estudios de Cohortes , Femenino , Pruebas Genéticas , Humanos , Riñón/metabolismo , Riñón/patología , Enfermedades Renales Quísticas/diagnóstico , Enfermedades Renales Quísticas/patología , Masculino , Fenotipo , Insuficiencia Renal Crónica/diagnóstico , Insuficiencia Renal Crónica/patología , Sistema Urinario/metabolismo , Sistema Urinario/patología , Secuenciación del ExomaRESUMEN
MicroRNAs (miRNAs) are endogenous non-coding RNAs that contain approximately 22 nucleotides. They serve as key regulators in various biological processes and their dysregulation is implicated in many diseases including cancer and autoimmune disorders. It has been well established that the maturation of miRNAs occurs in the cytoplasm and miRNAs exert post-transcriptional gene silencing (PTGS) via RNA-induced silencing complex (RISC) pathway in the cytoplasm. However, numerous studies reaffirm the existence of mature miRNA in the nucleus, and nucleus-cytoplasm transport mechanism has also been illustrated. Moreover, active regulatory functions of nuclear miRNAs were found including PTGS, transcriptional gene silencing (TGS), and transcriptional gene activation (TGA), in which miRNAs bind nascent RNA transcripts, gene promoter regions or enhancer regions and exert further effects via epigenetic pathways. Based on existing interaction rules, some miRNA binding sites prediction software tools are developed, which are evaluated in this article. In addition, we attempt to explore and review the nuclear functions of miRNA in immunity, tumorigenesis and invasiveness of tumor. As a non-canonical aspect of miRNA action, nuclear miRNAs supplement miRNA regulatory networks and could be applied in miRNA based therapies.
Asunto(s)
Núcleo Celular/genética , Regulación de la Expresión Génica , Inmunidad/genética , MicroARNs/genética , Neoplasias/genética , Interferencia de ARN , Animales , Biomarcadores , Núcleo Celular/metabolismo , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/inmunología , Transformación Celular Neoplásica/metabolismo , Biología Computacional/métodos , Citoplasma/genética , Citoplasma/metabolismo , Silenciador del Gen , Humanos , Neoplasias/metabolismo , Neoplasias/patología , Complejo Silenciador Inducido por ARN/metabolismo , Transducción de Señal , Programas InformáticosRESUMEN
Benefits of resistant starch (RS) consumption on host physiology encompass microbial activity-derived attenuation of intestinal inflammation. However, little is known about anti-inflammatory properties of RS of type 4. This study compared the effects of transglycosylated starch (TGS) consumption on the jejunal barrier function and expression of genes related to inflammation, barrier function and the mucosal defence in jejunum, ileum, caecum and colon of pigs. Moreover, interactions of TGS-induced alterations in bacterial metabolites and composition with host mucosal responses were assessed using sparse partial least squares regression and relevance network analysis. Intestinal samples were collected after pigs (n 8/diet; 4 months of age) were fed the experimental diets for 10 d. Consumption of TGS did not modify jejunal barrier function and gene expression. By contrast, TGS down-regulated the caecal expression of zonula occludens-1 and mucin 2 and of genes within the toll-like receptor 4 and NF-κB pro-inflammatory signalling cascade. Relevance networks revealed a microbiome signature on ileal, caecal and colonic mucosal signalling as TGS-derived changes in bacterial genera and fermentation acids, such as propionic acid, correlated with the differently expressed genes in ileum, caecum and colon of pigs. In conclusion, the present findings suggest certain anti-inflammatory capabilities of TGS by down-regulating the expression of pro-inflammatory pathways in the caecal mucosa, which seems to be mediated, at least in part, by TGS-induced changes in microbial action in the large intestine.