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1.
J Phys Chem Lett ; 11(3): 1141-1147, 2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-31951412

RESUMO

Double-electron electron resonance (DEER) can be used to track the structural dynamics of proteins in their native environment, the cell. This method provides the distance distribution between two spin labels attached at specific, well-defined positions in a protein. For the method to be viable under in-cell conditions, the spin label and its attachment to the protein should exhibit high chemical stability in the cell. Here we present low-temperature, trityl-trityl DEER distance measurements on two model proteins, PpiB (prolyl cis-trans isomerase from E. coli) and GB1 (immunoglobulin G-binding protein), doubly labeled with the trityl spin label, CT02MA. Both proteins gave in-cell distance distributions similar to those observed in vitro, with maxima at 4.5-5 nm, and the data were further compared with in-cell Gd(III)-Gd(III) DEER obtained for PpiB labeled with BrPSPy-DO3A-Gd(III) at the same positions. These results highlight the challenges of designing trityl tags suitable for in-cell distance determination at ambient temperatures on live cells.


Assuntos
Proteínas de Bactérias/química , Proteínas de Transporte/química , Ciclofilinas/química , Espectroscopia de Ressonância de Spin Eletrônica , Compostos de Tritil/química , Gadolínio/química , Marcadores de Spin
2.
Nat Commun ; 11(1): 575, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31996678

RESUMO

mTORC2 phosphorylates AKT in a hydrophobic motif site that is a biomarker of insulin sensitivity. In brown adipocytes, mTORC2 regulates glucose and lipid metabolism, however the mechanism has been unclear because downstream AKT signaling appears unaffected by mTORC2 loss. Here, by applying immunoblotting, targeted phosphoproteomics and metabolite profiling, we identify ATP-citrate lyase (ACLY) as a distinctly mTORC2-sensitive AKT substrate in brown preadipocytes. mTORC2 appears dispensable for most other AKT actions examined, indicating a previously unappreciated selectivity in mTORC2-AKT signaling. Rescue experiments suggest brown preadipocytes require the mTORC2/AKT/ACLY pathway to induce PPAR-gamma and establish the epigenetic landscape during differentiation. Evidence in mature brown adipocytes also suggests mTORC2 acts through ACLY to increase carbohydrate response element binding protein (ChREBP) activity, histone acetylation, and gluco-lipogenic gene expression. Substrate utilization studies additionally implicate mTORC2 in promoting acetyl-CoA synthesis from acetate through acetyl-CoA synthetase 2 (ACSS2). These data suggest that a principal mTORC2 action is controlling nuclear-cytoplasmic acetyl-CoA synthesis.


Assuntos
ATP Citrato (pro-S)-Liase/metabolismo , Adipócitos Marrons/metabolismo , Lipogênese/fisiologia , Alvo Mecanístico do Complexo 2 de Rapamicina/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Acetato-CoA Ligase/metabolismo , Animais , Proteínas de Transporte , Epigênese Genética , Ácido Graxo Sintases , Edição de Genes , Expressão Gênica , Transportador de Glucose Tipo 4/genética , Transportador de Glucose Tipo 4/metabolismo , Células HEK293 , Histonas/metabolismo , Humanos , Lipogênese/genética , Camundongos , Camundongos Endogâmicos C57BL , PPAR gama/metabolismo , Fosforilação , Proteômica , Elementos de Resposta
3.
Life Sci ; 240: 117117, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31790689

RESUMO

Adipocytokine alpha-2-glycoprotein 1 (AZGP1) is a 41-kDa protein which regulates insulin sensitivity and glycolipid metabolism. Recently, mounting evidence has indicated that AZGP1 plays a vital role in the progression and prognosis of many types of tumors, including hepatocellular carcinoma. Also, previous research has reported that AZGP1 levels are reduced significantly in patients with gastric carcinoma (GC). Here, we aim to assess the potential role and molecular mechanism underlying AZGP1-mediated regulation of GC progression. Both RT-PCR and Western blot methods demonstrated that AZGP1 levels were decreased in all GC cell lines tested, which included AGS, NCI-N87, MKN-28, SGC-7901 and MKN-45, relative to the normal human gastric mucosa epithelial (GES-1) cell line. Cell survival and proliferation rates were correspondingly were reduced, while cell apoptosis and caspase-3 activity were increased in NCI-N87 and SGC-7901 cells with high levels of AZGP1. Additionally, the mTOR signaling pathway was suppressed, whereas PTEN expression was elevated following transfection of NCI-N87 and SGC-7901 cells with an AZGP1 overexpressing plasmid. PTEN inhibition reversed the effects of AZGP1 on cell growth and apoptosis in SGC-7901 cells. Therefore, we conclude that AZGP1 induced apoptosis and growth inhibition in GC cells via the regulation of the mTOR/PTEN signaling pathway.


Assuntos
Apoptose/genética , Carcinoma/genética , Proteínas de Transporte/genética , Glicoproteínas/genética , PTEN Fosfo-Hidrolase/genética , Transdução de Sinais/genética , Neoplasias Gástricas/genética , Serina-Treonina Quinases TOR/genética , Proteínas de Transporte/biossíntese , Caspase 3/biossíntese , Caspase 3/genética , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular/genética , Glicoproteínas/biossíntese , Humanos
4.
Arch Insect Biochem Physiol ; 103(2): e21629, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31599031

RESUMO

Parasitoids serve as effective biocontrol agents for agricultural pests. However, they face constant challenges from host immune defense and numerous pathogens and must develop potent immune defense against these threats. Despite the recent advances in innate immunity, little is known about the immunological mechanisms of parasitoids. Here, we identified and characterized potential immune-related genes of the endoparasitoid, Pteromalus puparum, which act in regulating populations of some members of the Pieridae. We identified 216 immune-related genes based on interrogating the P. puparum genome and transcriptome databases. We categorized the cognate gene products into recognition molecules, signal moieties and effector proteins operating in four pathways, Toll, IMD, JAK/STAT, and JNK. Comparative analyses of immune-related genes from seven insect species indicate that recognition molecules and effector proteins are more expanded and diversified than signaling genes in these signal pathways. There are common 1:1 orthologs between the endoparasitoid P. puparum and its relative, the ectoparasitoid Nasonia vitripennis. The developmental expression profiles of immune genes randomly selected from the transcriptome analysis were verified by a quantitative polymerase chain reaction. Our work provides comprehensive analyses of P. puparum immune genes, some of which may be exploited in advancing parasitoid-based biocontrol technologies.


Assuntos
Proteínas de Transporte/genética , Imunidade Inata/genética , Proteínas de Insetos/genética , Transdução de Sinais/imunologia , Vespas/genética , Sequência de Aminoácidos , Animais , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Embrião não Mamífero/química , Embrião não Mamífero/metabolismo , Feminino , Perfilação da Expressão Gênica , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Larva/genética , Larva/metabolismo , Masculino , Filogenia , Pupa/genética , Pupa/metabolismo , Alinhamento de Sequência , Vespas/crescimento & desenvolvimento , Vespas/metabolismo , Vespas/fisiologia
5.
Gene ; 726: 144224, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-31669646

RESUMO

MicroRNA-155 (miR-155) has been identified to be overexpressed in various human cancers including osteosarcoma. However, whether the up-regulation of miR-155 is associated with osteosarcoma cancer stem cells (CSCs) is not well understood. In the present study, we showed that miR-155 induced the acquisition of stem-like features in U2OS osteosarcoma cells by increasing the expression of both CSCs surface markers (CD24, CD90, CD133) and CSC-related transcriptional factors (Nanog, SOX2, Oct4, Bim-1). Inflammatory factor TNF-α upregulated the miR-155 expression in U2OS cells and formed a feedback regulatory loop with miR-155. Furthermore, TNF-α/miR-155 axis promoted the cell proliferation, invasion and epithelial-mesenchymal transition (EMT) process in a TP53INP1 independent manner. We also revealed that TNF-α/miR-155 axis induced osteosarcoma CSCs transformation via ERK signaling pathway. These results indicate a crucial role of miR-155 in the acquisition of osteosarcoma CSC phenotype and miR-155 may serve as a potential target in future osteosarcoma therapy.


Assuntos
Proteínas de Transporte/genética , Transformação Celular Neoplásica/genética , Proteínas de Choque Térmico/genética , MicroRNAs/genética , Células-Tronco Neoplásicas/patologia , Osteossarcoma/genética , Fator de Necrose Tumoral alfa/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Transformação Celular Neoplásica/patologia , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Sistema de Sinalização das MAP Quinases/genética , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Osteossarcoma/patologia , Transdução de Sinais/genética , Fatores de Transcrição/genética
6.
Food Microbiol ; 86: 103325, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31703860

RESUMO

Clostridium perfringens SM101 genome encodes three serine proteases (CspA, CspB, and CspC), and genetic evidence indicates that CspB is required for processing of pro-SleC into active SleC, an enzyme essential for degradation of the peptidoglycan cortex during spore germination. In this study, the expression of cspA and cspC, as well as the germination and colony formation by spores of cspAC and cspC mutants of strain SM101, were assessed. We demonstrated that 1) the cspA and cspC genes were expressed as a bicistronic operon only during sporulation in the mother cell compartment of SM101; 2) both cspAC and cspC mutant spores were unable to germinate significantly with either KCl, l-glutamine, brain heart infusion (BHI) broth, or a 1:1 chelate of Ca2+ and dipicolinic acid (DPA); 3) consistent with germination results, both cspAC and cspC mutant spores were defective in normal DPA release; 4) the colony formation by cspAC and cspC mutant spores was ~106-fold lower than that of wild-type spores, although decoated mutant spores yielded wild-type level colony formation on plates containing lysozyme; 5) no processing of inactive pro-SleC into active SleC was observed in cspAC and cspC mutant spores during germination; and finally, 6) the defects in germination, DPA release, colony formation and SleC processing in cspAC and cspC mutant spores were complemented by the wild-type cspA-cspC operon. Collectively, these results indicate that both CspA and CspC are essential for C. perfringens spore germination through activating SleC and inducing cortex hydrolysis.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Transporte/metabolismo , Clostridium perfringens/enzimologia , Esporos Bacterianos/crescimento & desenvolvimento , Proteínas de Bactérias/genética , Proteínas de Transporte/genética , Clostridium perfringens/efeitos dos fármacos , Clostridium perfringens/genética , Clostridium perfringens/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Hidrólise , Óperon , Ácidos Picolínicos/farmacologia , Processamento de Proteína Pós-Traducional , Esporos Bacterianos/efeitos dos fármacos , Esporos Bacterianos/enzimologia , Esporos Bacterianos/genética
7.
Food Chem ; 308: 125576, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-31648092

RESUMO

This study investigated the effects of cold storage at different temperatures (4, -0.5, -3, and -20 °C) on protein degradation and its relationship to structural changes of black carp muscle. At -0.5 and 4 °C, major structural changes occurred, including the formation of gaps between myofibers and myofibrils, breakage of myofibrils and myofibers, and degradation of sarcoplasmic reticulum. Gel-based proteomic analysis showed that these structural changes were accompanied by degradation of a series of myofibrillar proteins, including titin, nebulin, troponin, myosin, myomesin, myosin-binding protein, and α-actinin. Loss of extractable gelatinolytic and caseinolytic protease activities was also observed. At -3 and -20 °C, formation of ice crystals was the most noticeable change. The major proteins were degraded at different locations in the black carp muscle, and gelatinolytic and caseinolytic proteases appear to contribute to the degradation of those proteins.


Assuntos
Carpas/metabolismo , Actinina/metabolismo , Animais , Proteínas de Transporte , Temperatura Baixa , Conectina/metabolismo , Cyprinidae/metabolismo , Proteínas Musculares/metabolismo , Miofibrilas/metabolismo , Miosinas/metabolismo , Proteólise , Proteômica
8.
Gene ; 725: 144163, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31639433

RESUMO

BACKGROUND: Previous studies have established that coronary artery disease is associated with excess inflammation. These studies have shown an elevation of both pro and anti-inflammatory cytokines in sufferers of coronary artery disease. There is increasing interest in the role played by the inflammasome Nod Like Receptor family pyrin domain containing 3 (NLRP3) in the aetiology of coronary artery disease. Increased severity of coronary artery disease correlates with higher levels of expression of NLRP3. Does NLRP3 polymorphisms play a role in the aetiology of coronary artery disease? METHOD: In a cohort of Vietnam War (n-299) veterans who have been previously exposed to trauma, NLRP3 polymorphisms were analysed for association with coronary calcium scores using analyses of variance. Independent t-test was used to analyse genotypes. In samples with a small representation of minor homozygotes, genotypes were combined and analysed using independent t-test. If any of the genotype analysis suggested the potential for a dominant or a recessive model the model was further explored. Hardy-Weinberg Equilibrium was calculated using Hardy-Weinberg equilibrium calculator including analysis for ascertainment bias. RESULTS: The NLRP3 polymorphism, rs10159239 was significantly associated (p = 0.001) with a higher raised coronary calcium score. The Single Nucleotide Polymorphism rs10159239 was examined by logistic regression with known risk factors for Coronary artery disease and remained significant (0.035). This is the first time rs10159239 A-allele has been associated with raised coronary calcium score. CONCLUSIONS: This is the first time rs10159239 A-allele has been associated with raised coronary calcium score. Further research is needed to replicate our results in larger well-characterised cohorts.


Assuntos
Doença da Artéria Coronariana/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Idoso , Idoso de 80 Anos ou mais , Alelos , Proteínas de Transporte/genética , Estudos de Coortes , Doença da Artéria Coronariana/metabolismo , Citocinas/genética , Grupo com Ancestrais do Continente Europeu/genética , Feminino , Frequência do Gene/genética , Predisposição Genética para Doença/genética , Genótipo , Humanos , Inflamação/genética , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Fatores de Risco , Veteranos , Guerra do Vietnã
9.
Biochemistry (Mosc) ; 84(12): 1490-1501, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31870253

RESUMO

The mammalian Golgi apparatus is a highly dynamic organelle, which is normally localized in the juxtanuclear space and plays an essential role in the regulation of cellular homeostasis. While posttranslational modification of cargo is mediated by the resident enzymes (glycosyltransferases, glycosidases, and kinases), the ribbon structure of Golgi and its cisternal stacking mostly rely on the cooperation of coiled-coil matrix golgins. Among them, giantin, GM130, and GRASPs are unique, because they form a tripartite complex and serve as Golgi docking sites for cargo delivered from the endoplasmic reticulum (ER). Golgi undergoes significant disorganization in many pathologies associated with a block of the ER-to-Golgi or intra-Golgi transport, including cancer, different neurological diseases, alcoholic liver damage, ischemic stress, viral infections, etc. In addition, Golgi fragments during apoptosis and mitosis. Here, we summarize and analyze clinically relevant observations indicating that Golgi fragmentation is associated with the selective loss of Golgi residency for some enzymes and, conversely, with the relocation of some cytoplasmic proteins to the Golgi. The central concept is that ER and Golgi stresses impair giantin docking site but have no impact on the GM130-GRASP65 complex, thus inducing mislocalization of giantin-sensitive enzymes only. This cardinally changes the processing of proteins by eliminating the pathways controlled by the missing enzymes and by activating the processes now driven by the GM130-GRASP65-dependent proteins. This type of Golgi disorganization is different from the one induced by the cytoskeleton alteration, which despite Golgi de-centralization, neither impairs function of golgins nor alters trafficking.


Assuntos
Complexo de Golgi/metabolismo , Animais , Autoantígenos/metabolismo , Sítios de Ligação , Proteínas de Transporte/metabolismo , Retículo Endoplasmático/metabolismo , Estresse do Retículo Endoplasmático , Humanos , Proteínas de Membrana/metabolismo , Neoplasias/metabolismo , Transporte Proteico
10.
Adv Exp Med Biol ; 1185: 97-101, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31884595

RESUMO

Leber congenital amaurosis (LCA) caused by AIPL1 mutations is one of the most severe forms of inherited retinal degeneration (IRD). The rapid and extensive photoreceptor degeneration challenges the development of potential treatments. Nevertheless, preclinical studies show that both gene augmentation and photoreceptor transplantation can regenerate and restore retinal function in animal models of AIPL1-associated LCA. However, questions regarding long-term benefit and safety still remain as these therapies advance towards clinical application. Ground-breaking advances in stem cell technology and genome editing are examples of alternative therapeutic approaches and address some of the limitations associated with previous methods. The continuous development of these cutting-edge biotechnologies paves the way towards a bright future not only for AIPL1-associated LCA patients but also other forms of IRD.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Terapia Baseada em Transplante de Células e Tecidos , Terapia Genética , Amaurose Congênita de Leber/terapia , Animais , Proteínas de Transporte , Proteínas do Olho/genética , Humanos , Amaurose Congênita de Leber/genética , Mutação
11.
Adv Exp Med Biol ; 1185: 531-535, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31884666

RESUMO

PRCD (progressive rod-cone degeneration) is a small ~6 kDa protein with unknown function that specifically resides in photoreceptor discs and interacts with rhodopsin. PRCD's discovery resulted from decades-long study of a canine retinal disease called progressive rod-cone degeneration which is one of the most frequent causes of blindness in dogs characterized by the slow, progressive death of rod photoreceptors followed by cones. A series of genetic studies eventually mapped the disease to a single point mutation in a novel gene which was then named Prcd. Highlighting the importance of this gene, this and several other mutations have been identified in human patients suffering from retinitis pigmentosa. In this review, we highlight what is currently known about PRCD protein, including the etiology and pathology of the retinal disease caused by its mutation, the protein's trafficking, localization, and biochemical characterization.


Assuntos
Proteínas de Transporte/genética , Proteínas do Olho/genética , Retinite Pigmentosa/genética , Animais , Cães , Humanos , Mutação , Células Fotorreceptoras Retinianas Cones/patologia , Células Fotorreceptoras Retinianas Bastonetes/patologia , Retinite Pigmentosa/patologia , Rodopsina/metabolismo
12.
Adv Exp Med Biol ; 1190: 23-31, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31760635

RESUMO

Regulation of differentiation and proliferation of Schwann cells is an essential part of the regulation of peripheral nerve development, degeneration, and regeneration. ZNRF1, a ubiquitin ligase, is expressed in undifferentiated/repair Schwann cells, directs glutamine synthetase to proteasomal degradation, and thereby increase glutamate levels in Schwann cell environment. Glutamate elicits subcellular signaling in Schwann cells via mGluR2 to modulate Neuregulin-1/ErbB2/3 signaling and thereby promote undifferentiated phenotype of Schwann cell.


Assuntos
Ácido Glutâmico/fisiologia , Bainha de Mielina/fisiologia , Nervos Periféricos/fisiologia , Transdução de Sinais , Proteínas de Transporte/fisiologia , Diferenciação Celular , Humanos , Receptores de AMPA/fisiologia , Células de Schwann
13.
Zhonghua Er Ke Za Zhi ; 57(11): 830-836, 2019 Nov 02.
Artigo em Chinês | MEDLINE | ID: mdl-31665836

RESUMO

Objective: To summarize the clinical and genetic features of ß-propeller protein-associated neurodegeneration (BPAN). Methods: The clinical data of 17 patients with BPAN with WDR45 gene variants were retrospectively collected at Children's Hospital of Fudan University, Peking University First Hospital, Capital Institute of Pediatrics, Shengjing Hospital of China Medical University and Shanghai Children's Hospital from June 2016 to December 2018, and their clinical manifestations, electroencephalogram, neuroimaging and genetics were analyzed. Results: Seventeen cases (13 females, 4 males), aged 1.1-8.8 years, were included. The median age of seizure onset was 14.5 months, from 3 months to 24 months of age, manifested with epileptic spasm in 6 cases and focal seizures in 5 cases. Eight patients had only one seizure type and 8 patients had two or more seizure types. Nine patients had complete remission of seizures. All 16 patients with seizures had developmental delay before the seizure onset, of whom 13 patients had moderate to severe seizures. The brain magnetic resonance imaging (MRI) was abnormal in 13 patients, including cerebral atrophy (10 cases) and thinning of the corpus callosum (9 cases). The brain magnetic susceptibility weighted imaging (SWI) in preschool stage showed prominent T2 hypointense signals in bilateral globus pallidus and brainstem ventral in two cases. Five seizure types (spasm, focal, absence, myodonic and generalized tonic clonic seizures)were found on ictal electroencephalogram(EEG) recordings. Compared to female patients(17(6-24) months of ege), male cases had earlier seizure onset (3, 4, 5, 18 months of age) . All patients had de novo variations in WDR45(6 nonsense, 4 frameshift, 3 missense and 4 splicing variations), with hemizygous variants in 3 males, mosaic variants in a male and heterozygous variants in 13 females, within which 5 variations had not been reported (c.977-1C>T,c.976+1G>C,c.10C>T,c.806del and c.110T>C). Conclusions: The patients with BPAN have profound developmental delay and are vulnerable to seizures. The male patients with BPAN tend to have more severer clinical phenotype than females. Early brain SWI could facilitate the timely diagnosis of this disease.


Assuntos
Proteínas de Transporte/genética , Epilepsia/genética , Doenças Neurodegenerativas/genética , China , Eletroencefalografia , Epilepsia/diagnóstico , Feminino , Humanos , Lactente , Masculino , Doenças Neurodegenerativas/diagnóstico por imagem , Estudos Retrospectivos , Convulsões
15.
Biomed Khim ; 65(5): 407-417, 2019 Aug.
Artigo em Russo | MEDLINE | ID: mdl-31666414

RESUMO

Isatin (indol-2,3-dione), an endogenous biofactor found in the brain, peripheral tissues and biological body fluids of humans and animals, exhibits a wide range of biological and pharmacological activities. They are realized via interaction with numerous isatin-binding proteins. Some of these proteins identified during proteomic profiling of the brain are involved in the development of neurodegenerative pathology. In the context of the neuroprotective effect, the effect of isatin is comparable to the effects of deprenyl (selegiline), a pharmacological agent used for treatment of Parkinson's disease. In this study, we have investigated the effect of a single dose administration of isatin (100 mg/kg) and deprenyl (10 mg/kg) to mice on the profile of the brain isatin-binding proteins. Comparative proteomic analysis of brain isatin-binding proteins of mice treated with isatin or deprenyl resulted in identification of a representative group of proteins (n=200) sensitive to the administration of these substances. The change in the profile of isatin-binding proteins may be obviously attributed to accumulation of isatin and deprenyl in the brain and their interaction with target proteins; this prevents protein binding to the affinity sorbent. Thus identified brain isatin-binding proteins of the control animals obviously represent specific targets that interact directly with isatin (and also with deprenyl) in vivo. Isatin or deprenyl administered to animals interact with these proteins and thus inhibit their binding to the affinity sorbent (immobilized isatin analogue).


Assuntos
Encéfalo/efeitos dos fármacos , Proteínas de Transporte/metabolismo , Isatina/farmacologia , Fármacos Neuroprotetores/farmacologia , Selegilina/farmacologia , Animais , Encéfalo/metabolismo , Camundongos , Proteômica
16.
Hum Genet ; 138(11-12): 1341-1357, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31667592

RESUMO

Recent research efforts to identify genes involved in malaria susceptibility using genome-wide approaches have focused on severe malaria. Here, we present the first GWAS on non-severe malaria designed to identify genetic variants involved in innate immunity or innate resistance mechanisms. Our study was performed on two cohorts of infants from southern Benin (525 and 250 individuals used as discovery and replication cohorts, respectively) closely followed from birth to 18-24 months of age, with an assessment of a space- and time-dependent environmental risk of exposure. Both the recurrence of mild malaria attacks and the recurrence of malaria infections as a whole (symptomatic and asymptomatic) were considered. Post-GWAS functional analyses were performed using positional, eQTL, and chromatin interaction mapping to identify the genes underlying association signals. Our study highlights a role of PTPRT, a tyrosine phosphatase receptor involved in STAT3 pathway, in the protection against both mild malaria attacks and malaria infections (p = 9.70 × 10-8 and p = 1.78 × 10-7, respectively, in the discovery cohort). Strong statistical support was also found for a role of MYLK4 (meta-analysis, p = 5.29 × 10-8 with malaria attacks), and for several other genes, whose biological functions are relevant in malaria infection. Results shows that GWAS on non-severe malaria can successfully identify new candidate genes and inform physiological mechanisms underlying natural protection against malaria.


Assuntos
Proteínas de Transporte/genética , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Malária/epidemiologia , Malária/genética , Locos de Características Quantitativas , Benin/epidemiologia , Pré-Escolar , Estudos de Coortes , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Malária/parasitologia , Masculino
17.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(11): 1081-1084, 2019 Nov 10.
Artigo em Chinês | MEDLINE | ID: mdl-31703130

RESUMO

OBJECTIVE: To detect pathogenic gene variants in two Chinese families with cone-rod dystrophy(CORD). METHODS: After the informed consent and comprehensive ophthalmic examinations for the patients, 3 mL peripheral blood was taken from the patients' blood vessel and DNA was extracted. The DNA was sequenced by whole-exome sequencing technology and variants were analyzed. RESULTS: Two novel compound heterozygous AIPL1 variants were detected in two patients, which were c.923T to C (p.L308P) and c.421C to T (p.Q141X) variants in Family 1, c.572T to C (p.L191P) and c.421C to T (p.Q141X) in Family 2. CONCLUSION: The results supported that AIPL1 gene variants are the main cause of the two CORD families. Whole-exome sequencing technology is a useful tool in the clinical differentiated diagnosis and genetic counseling for CORD patients.


Assuntos
Proteínas de Transporte/genética , Distrofias de Cones e Bastonetes/genética , Proteínas do Olho/genética , Proteínas Adaptadoras de Transdução de Sinal , Grupo com Ancestrais do Continente Asiático , Humanos , Mutação , Linhagem , Sequenciamento Completo do Exoma
18.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(11): 1115-1119, 2019 Nov 10.
Artigo em Chinês | MEDLINE | ID: mdl-31703139

RESUMO

OBJECTIVE: To explore the genetic basis for a boy with mental retardation. METHODS: Clinical data and peripheral blood samples of the family were collected. Potential variants were screened by using a panel for genes associated with intellectual impairment. Suspected variants were verified by PCR and Sanger sequencing. RESULTS: The child presented with mental retardation, language delay and poor self-care. Imaging analysis showed widening of brain fissures and subarachnoid space, and dysplasia of corpus callosum. Three novel heterozygous variants, namely c.1705T to C (p.S569P), c.1708dupC (p.R570Pfs*80) and c.2273delA (p.N758Tfs*22), were identified in the TRAPPC9 gene. The mother of the proband has carried the c.1708dupC (p.R570Pfs*80) and c.1705T to C (p.S569P) variants, while his father has carried the c.2273delA (p.N758Tfs*22) variant. CONCLUSION: The compound heterozygous variants of the TRAPPC9 gene probably underlie the disease in this family. Considering the clinical and genetic heterogeneity of mental retardation, genetic testing is essential for attaining diagnosis for patients with the relevant phenotype.


Assuntos
Proteínas de Transporte/genética , Deficiência Intelectual/genética , Criança , Testes Genéticos , Heterozigoto , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Masculino , Mutação , Fenótipo
19.
Infect Immun ; 88(1)2019 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-31636136

RESUMO

Streptococcus equi subsp. zooepidemicus is an important pathogen in horses that causes severe diseases such as pneumonia and abortion. Furthermore, it is a zoonotic agent, and contact with horses is a known risk factor. In this study, we investigated the working hypothesis that the zoonotic potential varies among S. equi subsp. zooepidemicus strains in association with differences in M-like protein-mediated binding of host plasma proteins. We demonstrate via in-frame deletion mutagenesis of two different S. equi subsp. zooepidemicus strains that the M-like protein SzM is crucial for the binding of fibrinogen to the bacterial surface and for survival in equine and human blood. S. equi subsp. zooepidemicus isolates of equine and human origins were compared with regard to SzM sequences and binding of equine and human fibrinogens. The N-terminal 216 amino acids of the mature SzM were found to exhibit a high degree of diversity, but the majority of human isolates grouped in three distinct SzM clusters. Plasma protein absorption assays and flow cytometry analysis revealed that pronounced binding of human fibrinogen is a common phenotype of human S. equi subsp. zooepidemicus isolates but much less so in equine S. equi subsp. zooepidemicus isolates. Furthermore, binding of human fibrinogen is associated with specific SzM types. These results suggest that SzM-mediated binding of human fibrinogen is an important virulence mechanism of zoonotic S. equi subsp. zooepidemicus isolates.


Assuntos
Antígenos de Bactérias/metabolismo , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Transporte/metabolismo , Fibrinogênio/metabolismo , Interações Hospedeiro-Patógeno , Streptococcus equi/metabolismo , Fatores de Virulência/metabolismo , Animais , Antígenos de Bactérias/classificação , Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/classificação , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Transporte/classificação , Proteínas de Transporte/genética , Análise por Conglomerados , Variação Genética , Cavalos , Humanos , Fenótipo , Ligação Proteica , Homologia de Sequência , Fatores de Virulência/classificação , Fatores de Virulência/genética
20.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 50(4): 540-545, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31642232

RESUMO

OBJECTIVE: To explore the anti-virus effect of AY358935 gene cloned by our research team on vesicular stomatitis virus (VSV), and studytheanti-virus mechanism. METHODS: HEK293 cells were stably transfected by the AY358935 gene recombinant plasmid pcDNA3.1-AY358935 or pcDNA3.1 blank plasmid respectively. Then VSV was added into the cell wells to infect the above cells at the multiplicity of infection (MOI) of 0.001. The virus titers in the liquid supernatant of the above three groups of cells were detected on different time, and the mortality of cells of each group was tested with trypan blue exclusion test at 24 h post VSV infection. Total RNA was extracted from the cells that stably transfected with target gene for the whole genome-wide cDNA microarray analysis. RESULTS: ① Virus titer:The virus titer in the liquid supernatant of pcDNA-3.1-AY358935 transfection cells group was obviously lower than those in pcDNA-3.1 transfection cell group and blank control cell group at 12 h post infection. The virus titerin the liquid supernatant of three groups were (7.16±2.33)×105 PFU/mL, (6.25±2.05)×106 PFU/mL and (7.75±2.54)×106 PFU/mL respectively at 18 h post infection. At that time, the virus titerin the liquid supernatant of pcDNA3.1-AY358935 group was nearly 10 times lower than those of other two groups (P < 0.01). ②Mortality of cells:The cell mortality of pcDNA3.1-AY358935 group, pcDNA3.1 group and blank group were (35.00±6.68)%, (78.33±15.03)% and (83.34±14.98)% respectively at 24 h post infection.The cell mortality of pcDNA3.1-AY358935 group was significantly decreased comparing with other two groups (P < 0.01). ③Result of genes chip analysis: compared with pcDNA3.1 group, 30 cell genes were up-regulated by more than 3 times in pcDNA3.1-AY358935 group. Among them, the proportion of interferon-activating gene, interferon-effect gene, cytokine and chemokine was 27%, 17%, and 20%, respectively. CONCLUSION: AY358935 gene hasan anti-VSV effect, and its anti-virus mechanism may involve the interferon-associated natural immune response.


Assuntos
Proteínas de Transporte/genética , Estomatite Vesicular/imunologia , Animais , Citocinas , Células HEK293 , Humanos , Interferons , Plasmídeos , Transfecção , Vesiculovirus
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