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1.
Zhonghua Zhong Liu Za Zhi ; 41(11): 826-830, 2019 Nov 23.
Artigo em Chinês | MEDLINE | ID: mdl-31770849

RESUMO

Objective: To investigate the expression of fragile-site associated tumor suppressor (FATS) in non-small cell lung cancer and its relationship with clinicopathological features and prognosis. Methods: A total of 140 non-small cell lung cancer (NSCLC) cases and 30 adjacent normal tissues were used to detect the expression level of FATS protein, and to analyze the relationship of FATS protein expression and clinicopathological features and prognosis of NSCLC. Results: Western blot showed that the expression of FATS in adjacent normal tissues was significantly higher than that in non-small cell lung cancer tissues. The results of immunohistochemistry showed that the high expression rate of FATS in 140 cases of NSCLC was 40.0%, and the high expression rate of FATS in 30 cases of adjacent tissues was 73.3%. The difference was statistically significant (P=0.01). Further analysis showed that the TNM stage (P=0.044) and lymph node metastasis (P=0.022) were significant difference between FATS high expression group and low expression group. The 6-year overall survival (OS) rates of NSCLC patients with FATS high-expression and low-expression were 57.1% and 23.8%, respectively, and the 6-year disease-free survival (DFS) rates were 53.6% and 21.4%, respectively, with statistically significant differences (P=0.001). In Cox multivariate analysis, we found gender (HR=1.658, P=0.028; HR=1.684, P=0.023), TNM staging (HR=2.327, P=0.019; HR=2.332, P=0.013) and FATS expression (HR=0.532, P=0.010; HR=0.538, P=0.009) were independent prognostic factors for both OS and DFS of NSCLC patients. Conclusions: The expression of FATS protein is associated with the development and is an independent prognostic factor of NSCLC patients. The detection of FATS protein is expected to be a new biomarker for evaluating the prognosis of NSCLC patients.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Proteínas do Citoesqueleto/genética , Neoplasias Pulmonares/genética , Biomarcadores Tumorais/genética , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Humanos , Estimativa de Kaplan-Meier , Neoplasias Pulmonares/diagnóstico , Estadiamento de Neoplasias , Prognóstico
2.
Nat Cell Biol ; 21(10): 1191-1205, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31548610

RESUMO

Cells of multicellular organisms need to adopt specific morphologies. However, the molecular mechanisms bringing about membrane topology changes are far from understood-mainly because knowledge of membrane-shaping proteins that can promote local membrane curvatures is still limited. Our analyses unveiled that several members of a large, previously unrecognised protein family, which we termed N-Ank proteins, use a combination of their ankyrin repeat array and an amino (N)-terminal amphipathic helix to bind and shape membranes. Consistently, functional analyses revealed that the N-Ank protein ankycorbin (NORPEG/RAI14), which was exemplarily characterised further, plays an important, ankyrin repeat-based and N-terminal amphipathic helix-dependent role in early morphogenesis of neurons. This function furthermore required coiled coil-mediated self-assembly and manifested as ankycorbin nanodomains marked by protrusive membrane topologies. In summary, here, we unveil a class of powerful membrane shapers and thereby assign mechanistic and cell biological functions to the N-Ank protein superfamily.


Assuntos
Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Proteínas do Citoesqueleto/química , Proteínas do Citoesqueleto/metabolismo , Morfogênese , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo , Animais , Repetição de Anquirina/genética , Células Cultivadas , Proteínas do Citoesqueleto/genética , Células HEK293 , Células HeLa , Humanos , Camundongos , Modelos Moleculares , Neurônios/citologia , Neurônios/metabolismo , Domínios Proteicos/genética , Ratos , Fatores de Transcrição/genética
3.
DNA Cell Biol ; 38(11): 1323-1337, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31536386

RESUMO

Our previous study has indicated that the parathyroid hormone type 1 receptor (PTHR1) may play important roles in development and progression of osteosarcoma (OS) by regulating Wnt, angiogenesis, and inflammation pathway genes. The goal of this study was to further illuminate the roles of PTHR1 in OS by investigating upstream regulation mechanisms (including microRNA [miRNA] and transcription factors [TFs]) of crucial genes. The microarray dataset GSE46861 was downloaded from the Gene Expression Omnibus database, in which six tumors with short hairpin RNA (shRNA) PTHR1 knockdown (PTHR1.358) and six tumors with shRNA control knockdown (Ren.1309) were collected from mice. Differentially expressed genes (DEGs) between PTHR1.358 and Ren.1309 were identified using the linear models for microarray data (LIMMA) method, and then the miRNA-TF-mRNA regulatory network was constructed using data from corresponding databases, followed by module analysis, to screen crucial regulatory relationships. OS-related human miRNAs were extracted from the curated Osteosarcoma Database. Gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were enriched using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) tool. As a result, the miRNA-TF-mRNA regulatory network, including 1049 nodes (516 miRNA, 25 TFs, and 508 DEGs) and 15942 edges (interaction relationships, such as Pparg-Abca1 and miR-590-3p-AXIN2), was constructed, from which three significant modules were extracted and modules 2 and 3 contained interactions between miRNAs/TFs and DEGs such as miR-103-3p-AXIN2, miR-124-3p-AR-Tgfb1i1, and miR-27a-3p-PPARG-Abca1. miR-27a-3p was a known miRNA associated with OS. Abca1, AR, and miR-124-3p were hub genes in the miRNA-TF-mRNA network. Tgfb1i1 was involved in cell proliferation, Abca1 participated in the cholesterol metabolic process, and AXIN2 was associated with the canonical Wnt signaling pathway. Furthermore, we also confirmed upregulation of miR-590-3p and downregulation of AXIN2 in the mouse OS cell line K7M2-WT transfected with PTHR1 shRNA. In conclusion, PTHR1 may play important roles in progression of OS by activating miR-124-3p-AR-Tgfb1i1, miR-27a-3p-PPARG-Abca1, and miR-103/590-3p-AXIN2 axes.


Assuntos
Neoplasias Ósseas , Osteossarcoma , Receptor Tipo 1 de Hormônio Paratireóideo/fisiologia , Transportador 1 de Cassete de Ligação de ATP/genética , Transportador 1 de Cassete de Ligação de ATP/fisiologia , Animais , Proteína Axina/genética , Proteína Axina/fisiologia , Neoplasias Ósseas/genética , Neoplasias Ósseas/patologia , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/fisiologia , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/fisiologia , Progressão da Doença , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Células HEK293 , Humanos , Proteínas com Domínio LIM/genética , Proteínas com Domínio LIM/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , MicroRNAs/fisiologia , Osteossarcoma/genética , Osteossarcoma/patologia , PPAR gama/genética , PPAR gama/fisiologia , Receptor Tipo 1 de Hormônio Paratireóideo/genética , Receptores Androgênicos/genética , Receptores Androgênicos/fisiologia , Transdução de Sinais/genética , Células Tumorais Cultivadas
4.
Microbiol Res ; 228: 126304, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31422235

RESUMO

Streptococcus suis (S. suis) is an important zoonotic pathogen that causes major economic losses in the pig industry worldwide. The S. suis cell division process is an integral part of its growth and reproduction, which is controlled by a complex regulatory network. Pyruvate dehydrogenase (PDH), which catalyzes the oxidative decarboxylation of pyruvate to form acetyl-CoA, while reducing NAD + to NADH, plays an important role in energy metabolism. Recently, we reported that pdh regulates virulence by reducing stress tolerance and biofilm formation in S. suis serotype 2. In this study, we found that deletion of the pdh gene in S. suis resulted in abnormal cell chains, plump morphology and abnormal localization of the Z rings, indicating that the knockout mutant is impaired in its ability to divide. In addition, the interaction between FtsZ and PDH in vitro was confirmed by ELISA, and qRT-PCR analysis revealed that the deletion of the pdh gene results in differential expression of the division-related genes ftsZ, ftsK, ftsl, zapA, divIC, pbp1a, rodA, mreD, and sepF. These results indicate that pdh is involved in the normal formation of Z rings and cell morphology during S. suis cell division.


Assuntos
Divisão Celular/genética , Divisão Celular/fisiologia , Complexo Piruvato Desidrogenase/genética , Streptococcus suis/citologia , Streptococcus suis/genética , Streptococcus suis/fisiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Proteínas do Citoesqueleto/genética , Deleção de Genes , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Técnicas de Inativação de Genes , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/veterinária , Streptococcus suis/patogenicidade , Suínos , Virulência , Fatores de Virulência/genética
5.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(7): 662-665, 2019 Jul 10.
Artigo em Chinês | MEDLINE | ID: mdl-31302906

RESUMO

OBJECTIVE: To screen for MYOC gene variants among sporadic patients with primary open angle glaucoma (POAG). METHODS: For 398 patients with POAG, Sanger sequencing was applied to detect potential variants of the MYOC gene. RESULTS: Eight patients (2.0%) were found to harbor variations of the MYOC gene. These included five types of variants, among which c.667C>T (p.Pro223Ser) and c.1138G>T (p.Asp380Tyr) were novel. c.382C>T (p.Arg128Trp), c.1109C>T(p.Pro370Leu) and c.1130C>A (p.Thr377Lys) were previously associated with POAG. Alignment of amino acid sequences of MYOC proteins of various species revealed that the two novel variants have occurred at highly conserved positions. c.1138G>T was predicted to be possible pathogenic by Bioinformatic analysis. CONCLUSION: Two novel variants of the MYOC gene were detected among sporadic POAG patients, which enriched its variant spectrum.


Assuntos
Proteínas do Citoesqueleto/genética , Proteínas do Olho/genética , Glaucoma de Ângulo Aberto/genética , Glicoproteínas/genética , Humanos , Mutação
6.
Cell Mol Biol Lett ; 24: 48, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31333725

RESUMO

Background: In recent years, microRNA-211 (miR211) has been considered as a tumor suppressor in multiple malignancies. However, the function of miR211 in human osteosarcoma has not been explored intensively so far. In this study, the relationship between miR211 and EZRIN was analyzed in human osteosarcoma. Methods: The expression levels of miR211 and EZRIN were measured in both human osteosarcoma cells and tissues. The direct regulatory relationship between miR211 and EZRIN was evaluated using dual-luciferase assay. The effect of miR211 and EZRIN overexpression on cell proliferation, migration/invasion, and apoptosis was detected. Results: The expression of miR211 was obviously lower in osteosarcoma tissues than paracancerous tissues. EZRIN was identified as the direct target of miR211, and up-regulation of miR211 increased the percentage of cell apoptosis, and suppressed cell proliferation as well as cell migration/invasion via directly regulating EZRIN. Conclusions: Our study indicated that miR211 has an important role in the development and progress of osteosarcoma, and it might become a novel target in the diagnosis and treatment of human osteosarcoma.


Assuntos
Apoptose , Neoplasias Ósseas/metabolismo , Proliferação de Células , Proteínas do Citoesqueleto/genética , MicroRNAs/metabolismo , Osteossarcoma/metabolismo , Neoplasias Ósseas/fisiopatologia , Linhagem Celular Tumoral , Movimento Celular , Regulação Neoplásica da Expressão Gênica , Genes Supressores de Tumor , Humanos , MicroRNAs/genética , Osteossarcoma/fisiopatologia
7.
Int J Mol Sci ; 20(11)2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-31151140

RESUMO

As a conserved actin-regulating protein, CAP (adenylyl Cyclase-Associated Protein) functions to facilitate the rearrangement of the actin cytoskeleton. The ubiquitously expressed isoform CAP1 drives mammalian cell migration, and accordingly, most studies on the involvement of CAP1 in human cancers have largely been based on the rationale that up-regulated CAP1 will stimulate cancer cell migration and invasiveness. While findings from some studies reported so far support this case, lines of evidence largely from our recent studies point to a more complex and profound role for CAP1 in the invasiveness of cancer cells, where the potential activation of cell adhesion signaling is believed to play a key role. Moreover, CAP1 was also found to control proliferation in breast cancer cells, through the regulation of ERK (External signal-Regulated Kinase). Alterations in the activities of FAK (Focal Adhesion Kinase) and ERK from CAP1 depletion that are consistent to the opposite adhesion and proliferation phenotypes were detected in the metastatic and non-metastatic breast cancer cells. In this review, we begin with the overview of the literature on CAP, by highlighting the molecular functions of mammalian CAP1 in regulating the actin cytoskeleton and cell adhesion. We will next discuss the role of the FAK/ERK axis, and possibly Rap1, in mediating CAP1 signals to control breast cancer cell adhesion, invasiveness, and proliferation, largely based on our latest findings. Finally, we will discuss the relevance of these novel mechanistic insights to ultimately realizing the translational potential of CAP1 in targeted therapeutics for breast cancer.


Assuntos
Neoplasias da Mama/metabolismo , Proteínas de Ciclo Celular/metabolismo , Proteínas do Citoesqueleto/metabolismo , Transdução de Sinais , Citoesqueleto de Actina/metabolismo , Animais , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Adesão Celular/genética , Proteínas de Ciclo Celular/química , Proteínas de Ciclo Celular/genética , Movimento Celular/genética , Proliferação de Células , Proteínas do Citoesqueleto/química , Proteínas do Citoesqueleto/genética , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Humanos , Ligação Proteica , Relação Estrutura-Atividade , Pesquisa Médica Translacional
8.
Plant Sci ; 285: 99-109, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31203898

RESUMO

Seed development is a complex regulatory process that includes a transition from gametophytic to sporophytic program. The synchronized development of different seed compartments (seed coat, endosperm and embryo) depends on a balance in parental genome contributions. In the most severe cases, the disruption of the balance leads to seed abortion. This represents one of the main obstacles for the engineering of asexual reproduction through seeds (apomixis), and for generating new interspecies hybrids. The repression of auxin synthesis by the Polycomb Repressive Complex 2 (PRC2) is a major mechanism contributing to sensing genome balance. However, current efforts focusing on decreasing PRC2 or elevating auxin levels have not yet resulted in the production of apomictic seed. Here, we show that EMSY-Like Tudor/Agenet H3K36me3 histone readers EML1 and EML3 are necessary for early stages of seed development to proceed at a normal rate in Arabidopsis. We further demonstrate that both EML1 and EML3 are required to prevent the initiation of seed development in the absence of fertilization. Based on the whole genome expression analysis, we identify auxin transport and signaling genes as the most enriched downstream targets of EML1 and EML3. We hypothesize that EML1 and EML3 function to repress and soften paternal gene expression by fine-tuning auxin responses. Discovery of this pathway may contribute to the engineering of apomixis and interspecies hybrids.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/crescimento & desenvolvimento , Proteínas do Citoesqueleto/fisiologia , Histonas/metabolismo , Proteínas Nucleares/fisiologia , Sementes/crescimento & desenvolvimento , Apomixia , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Proteínas do Citoesqueleto/genética , Fertilização , Proteínas Nucleares/genética , Filogenia , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase , Sementes/fisiologia
9.
Med Sci Monit ; 25: 3374-3389, 2019 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-31063460

RESUMO

BACKGROUND The protein 4.1 family is a family of cytoskeletal proteins that play an important role in maintaining normal cell morphology and cell adhesion, migration, division, and intercellular signaling. The main aim of this study was to explore the prognostic significance of the protein 4.1 family in breast cancer (BC) patients and to provide new biomarkers and therapeutic targets for the diagnosis and treatment of BC. MATERIAL AND METHODS The expression of 4.1 family members in various tumor types was compared to normal controls using the ONCOMINE and GOBO databases. The prognostic significance of the 4.1 family in BC patients was determined by Kaplan-Meier Plotter. RESULTS EPB41L2 (4.1G) was expressed at higher levels in normal tissues compared with BC patients for all 4.1 family members. In survival analysis, 4.1G and EPB41 (4.1R) mRNA high expressions were associated with better survival in BC patients. Moreover, 4.1G high expression was significantly associated with longer overall survival (OS) in luminal A and protracted relapse-free survival (RFS) in luminal B subtype BC patients who received Tamoxifen treatment. In addition, high expression of each 4.1 family member also showed better prognostic value in different molecular subtypes of BC. CONCLUSIONS These results indicate that the protein 4.1 family can be regarded as novel biomarkers and potential therapeutic targets for BC. Further research is needed to explore the detailed biological functions.


Assuntos
Neoplasias da Mama/patologia , Proteínas do Citoesqueleto/biossíntese , Proteínas de Membrana/biossíntese , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Proteínas do Citoesqueleto/genética , Bases de Dados Genéticas , Intervalo Livre de Doença , Feminino , Humanos , Estimativa de Kaplan-Meier , Proteínas de Membrana/genética , Prognóstico , Análise de Sobrevida
10.
Orphanet J Rare Dis ; 14(1): 114, 2019 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-31122244

RESUMO

BACKGROUND: Current diagnostic tests for hereditary spherocytosis (HS) focus on the detection of hemolysis or indirectly assessing defects of membrane protein, whereas direct methods to detect protein defects are complicated and difficult to implement. In the present study, we investigated the patterns of genetic variation associated with HS among patients clinically diagnosed with HS. METHODS: Multi-gene targeted sequencing of 43 genes (17 RBC membrane protein-encoding genes, 20 RBC enzyme-encoding genes, and six additional genes for the differential diagnosis) was performed using the Illumina HiSeq platform. RESULTS: Among 59 patients with HS, 50 (84.7%) had one or more significant variants in a RBC membrane protein-encoding genes. A total of 54 significant variants including 46 novel mutations were detected in six RBC membrane protein-encoding genes, with the highest number of variants found in SPTB (n = 28), and followed by ANK1 (n = 19), SLC4A1 (n = 3), SPTA1 (n = 2), EPB41 (n = 1), and EPB42 (n = 1). Concurrent mutations of genes encoding RBC enzymes (ALDOB, GAPDH, and GSR) were detected in three patients. UGT1A1 mutations were present in 24 patients (40.7%). Positive rate of osmotic fragility test was 86.8% among patients harboring HS-related gene mutations. CONCLUSIONS: This constitutes the first large-scaled genetic study of Korean patients with HS. We demonstrated that multi-gene target sequencing is sensitive and feasible that can be used as a powerful tool for diagnosing HS. Considering the discrepancies of clinical and molecular diagnoses of HS, our findings suggest that molecular genetic analysis is required for accurate diagnosis of HS.


Assuntos
Fragilidade Osmótica/fisiologia , Esferócitos/metabolismo , Esferocitose Hereditária/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteína 1 de Troca de Ânion do Eritrócito/genética , Proteína 1 de Troca de Ânion do Eritrócito/metabolismo , Anquirinas/genética , Anquirinas/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Criança , Pré-Escolar , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Feminino , Glucuronosiltransferase/genética , Glucuronosiltransferase/metabolismo , Humanos , Lactente , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Pessoa de Meia-Idade , Mutação/genética , Fragilidade Osmótica/genética , Patologia Molecular , República da Coreia , Espectrina/genética , Espectrina/metabolismo , Esferocitose Hereditária/genética , Adulto Jovem
11.
PLoS Genet ; 15(5): e1008161, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31107865

RESUMO

During early meiotic prophase, homologous chromosomes are connected along their entire lengths by a proteinaceous tripartite structure known as the synaptonemal complex (SC). Although the components that comprise the SC are predominantly studied in this canonical ribbon-like structure, they can also polymerize into repeated structures known as polycomplexes. We find that in Drosophila oocytes, the ability of SC components to assemble into canonical tripartite SC requires the E3 ubiquitin ligase Seven in absentia (Sina). In sina mutant oocytes, SC components assemble into large rod-like polycomplexes instead of proper SC. Thus, the wild-type Sina protein inhibits the polymerization of SC components, including those of the lateral element, into polycomplexes. These polycomplexes persist into meiotic stages when canonical SC has been disassembled, indicating that Sina also plays a role in controlling SC disassembly. Polycomplexes induced by loss-of-function sina mutations associate with centromeres, sites of double-strand breaks, and cohesins. Perhaps as a consequence of these associations, centromere clustering is defective and crossing over is reduced. These results suggest that while features of the polycomplexes can be recognized as SC by other components of the meiotic nucleus, polycomplexes nonetheless fail to execute core functions of canonical SC.


Assuntos
Proteínas Nucleares/metabolismo , Proteínas Nucleares/fisiologia , Complexo Sinaptonêmico/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/fisiologia , Animais , Proteínas de Ciclo Celular/metabolismo , Núcleo Celular/metabolismo , Centrômero/metabolismo , Proteínas Cromossômicas não Histona/metabolismo , Pareamento Cromossômico/genética , Proteínas do Citoesqueleto/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Feminino , Meiose , Oócitos/metabolismo , Complexo Sinaptonêmico/genética
12.
Parasit Vectors ; 12(1): 227, 2019 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-31088539

RESUMO

BACKGROUND: Giardia lamblia, a protozoan pathogen causing diarrheal outbreaks, has characteristic cytoskeletal structures including eight flagella, a median body and a ventral disc. Gamma-giardin is a unique component protein of the cytoskeleton of this protozoan. RESULTS: Through comparative proteomic analysis between different stages of the cell cycle, G. lamblia γ-giardin (Glγ-giardin) was identified as an upregulated protein in the G2-phase. Increased Glγ-giardin expression in G2 was confirmed by western blot and real-time polymerase chain reaction analyses. Knockdown of this protein using a morpholino affected the formation of ventral discs, especially the microribbons of the discs, but exerted little effect on the binding ability of G. lamblia. The number of cells with four nuclei was increased in Glγ-giardin-knockdown cells. Expression of Glγ-giardin was decreased during encystation, in contrast with the G2-phase. CONCLUSIONS: Knockdown experiments demonstrated that Glγ-giardin is a component of the trilaminar structure of the ventral disc. Expression of Glγ-giardin is induced in the G2-phase prior to active cell division, whereas its expression decreases during encystation, a dormant stage of G. lamblia.


Assuntos
Proteínas do Citoesqueleto/genética , Fase G2/genética , Giardia lamblia/citologia , Giardia lamblia/crescimento & desenvolvimento , Proteínas de Protozoários/genética , Proteínas do Citoesqueleto/metabolismo , Citoesqueleto/química , Citoesqueleto/genética , Técnicas de Silenciamento de Genes , Encistamento de Parasitas , Proteômica , Proteínas de Protozoários/metabolismo , Alinhamento de Sequência , Regulação para Cima
13.
Immunity ; 50(6): 1412-1424.e6, 2019 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-31076360

RESUMO

Assembly of inflammasomes after infection or injury leads to the release of interleukin-1ß (IL-1ß) and to pyroptosis. After inflammasome activation, cells either pyroptose or enter a hyperactivated state defined by IL-1ß secretion without cell death, but what controls these different outcomes is unknown. Here, we show that removal of the Toll-IL-1R protein SARM from macrophages uncouples inflammasome-dependent cytokine release and pyroptosis, whereby cells displayed increased IL-1ß production but reduced pyroptosis. Correspondingly, increasing SARM in cells caused less IL-1ß release and more pyroptosis. SARM suppressed IL-1ß by directly restraining the NLRP3 inflammasome and, hence, caspase-1 activation. Consistent with a role for SARM in pyroptosis, Sarm1-/- mice were protected from lipopolysaccharide (LPS)-stimulated sepsis. Pyroptosis-inducing, but not hyperactivating, NLRP3 stimulants caused SARM-dependent mitochondrial depolarization. Thus, SARM-dependent mitochondrial depolarization distinguishes NLRP3 activators that cause pyroptosis from those that do not, and SARM modulation represents a cell-intrinsic mechanism to regulate cell fate after inflammasome activation.


Assuntos
Proteínas do Domínio Armadillo/metabolismo , Citocinas/metabolismo , Proteínas do Citoesqueleto/metabolismo , Inflamassomos/metabolismo , Animais , Proteínas do Domínio Armadillo/genética , Biomarcadores , Sobrevivência Celular , Proteínas do Citoesqueleto/genética , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Camundongos Knockout , Mitocôndrias/genética , Mitocôndrias/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Ligação Proteica , Piroptose , Transdução de Sinais
14.
Acta Crystallogr F Struct Biol Commun ; 75(Pt 5): 359-367, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31045565

RESUMO

As of 2017, tuberculosis had infected 1.7 billion people (23% of the population of the world) and caused ten million deaths. Mycobacterium tuberculosis (Mtb) is quickly evolving, and new strains are classified as multidrug resistant. Thus, the identification of novel druggable targets is essential to combat the proliferation of these drug-resistant strains. Filamenting temperature-sensitive mutant Z (FtsZ) is a key protein involved in cytokinesis, an important process for Mtb proliferation and viability. FtsZ is required for bacterial cell division because it polymerizes into a structure called the Z-ring, which recruits accessory division proteins to the septum. Here, the crystal structure of the MtbFtsZ protein has been determined to 3.46 Šresolution and is described as a dimer of trimers, with an inter-subunit interface between protomers AB and DE. In this work, a novel conformation of MtbFtsZ is revealed involving the T9 loop and the nucleotide-binding pocket of protomers BC and EF.


Assuntos
Proteínas de Bactérias/química , Proteínas do Citoesqueleto/química , Mycobacterium tuberculosis/química , Subunidades Proteicas/química , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Divisão Celular , Clonagem Molecular , Cristalografia por Raios X , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Cinética , Modelos Moleculares , Mutação , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Multimerização Proteica , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homologia Estrutural de Proteína , Temperatura Ambiente
15.
Nat Commun ; 10(1): 1901, 2019 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-31015409

RESUMO

Asymmetric cell division is a major mechanism generating cell diversity. As cell cycle duration varies among cells in mammalian tissue culture cells, we asked whether their division asymmetry contributes to this variability. We identify among sibling cells an outlier using hierarchical clustering on cell cycle durations of granddaughter cells obtained by lineage tracking of single histone2B-labelled MDCKs. Remarkably, divisions involving outlier cells are not uniformly distributed in lineages, as shown by permutation tests, but appear to emerge from asymmetric divisions taking place at non-stochastic levels: a parent cell influences with 95% confidence and 0.5% error the unequal partitioning of the cell cycle duration in its two progenies. Upon ninein downregulation, this variability propagation is lost, and outlier frequency and variability in cell cycle durations in lineages is reduced. As external influences are not detectable, we propose that a cell-autonomous process, possibly involved in cell specialisation, determines cell cycle duration variability.


Assuntos
Divisão Celular Assimétrica , Linhagem da Célula/genética , Proteínas do Citoesqueleto/genética , Escherichia coli/citologia , Histonas/genética , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Rastreamento de Células/métodos , Proteínas do Citoesqueleto/metabolismo , Cães , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Genes Reporter , Histonas/metabolismo , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Células Madin Darby de Rim Canino , Modelos Biológicos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Fatores de Tempo
16.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(5): 447-450, 2019 May 10.
Artigo em Chinês | MEDLINE | ID: mdl-31030430

RESUMO

OBJECTIVE: To explore the genetic etiology of two pedigrees affected with congenital arthrogryposis. METHODS: Whole exome sequencing (WES) was used to screen potential variations in the proband. Suspected variations were analyzed with bioinformatics software and validated by Sanger sequencing. RESULTS: A heterozygous c.1123G>A (p.Glu375Lys) variation was detected in the proband and an affected fetus from pedigree 1, while a de novo heterozygous c.118 G>A (p.Val40Met) variation was detected in an affected fetus from pedigree 2. CONCLUSION: The two heterozygous variations of the MYH3 gene probably underlie the disease in the pedigrees. Above results have facilitated genetic counseling and prenatal diagnosis.


Assuntos
Artrogripose , Proteínas do Citoesqueleto/genética , Feminino , Heterozigoto , Humanos , Mutação , Linhagem , Gravidez , Diagnóstico Pré-Natal , Sequenciamento Completo do Exoma
17.
Vet Parasitol ; 268: 32-35, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30981303

RESUMO

This study developed and evaluated Giardia duodenalis cyst propagation using a dexamethasone immunosuppressed CF-1 mouse model as an alternative to a previously described Mongolian gerbil model. The CF-1 mouse model shed significantly more cysts per animal during a 16-18 h collection period compared to the gerbil (averages: 7.8 × 106 cysts/CF-1 mouse and 2.5 × 106 cysts/gerbil). In addition, the patency period for this model differed from both G. muris in mice and G. duodenalis in gerbils in that cysts were shed continuously for over 20 days. Results further showed that the ß-giardin gene sequences from gerbil derived and mouse derived G. duodenalis were identical, after 34 serial passages through the CF-1 mouse model. Overall, the CF-1 mouse model produced higher concentrations of cysts per animal, and were genetically and phenotypically stable based on ß-giardin gene sequences.


Assuntos
Cistos/parasitologia , Modelos Animais de Doenças , Giardia lamblia/crescimento & desenvolvimento , Hospedeiro Imunocomprometido , Animais , Anti-Inflamatórios/administração & dosagem , Proteínas do Citoesqueleto/genética , Dexametasona/administração & dosagem , Fezes/parasitologia , Feminino , Genótipo , Giardia lamblia/genética , Giardíase/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos , Proteínas de Protozoários/genética , Reprodução
18.
Eur J Pharm Sci ; 135: 103-112, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31034983

RESUMO

Tuberculosis, caused by Mycobacterium tuberculosis has been one of the primal afflictions to human, and owing to the current scenario of drug resistance, newer drugs, and alternate targets are required to mitigate the disease. FtsZ is a GTP hydrolyzing protein, conserved in prokaryotes that plays a central role in Z-ring formation during cell division cytokinesis stage. This study employs the use of pharmacophore models derived from two different datasets based on Mtb-FtsZ GTPase inhibition and whole cell antibacterial activity, to virtually screen and shortlist novel compounds from In-house small molecule library as Mtb-FtsZ inhibitors and evaluate their in-vitro and ex-vivo activity. The results revealed Piperine (IC50 = 21.2 ±â€¯0.7 µM), 4-Bromo di-methoxy coumarin (IC50 = 13.0 ±â€¯1.6 µM) and Di-ethyl amino methyl coumarin (IC50 = 19.4 ±â€¯1.1) as potent Mtb-FtsZ GTPase inhibitors which showed considerable antibacterial activity (84.0 ±â€¯2.6 µM, 56.0 ±â€¯4.3 µM and 108 ±â€¯7.1 µM respectively) against M. smegmatis. They appear to be bacteriostatic, as well as treatment with these compounds led to a 3× increase in cell length of M. smegmatis. Further these molecules also altered the FtsZ gene expression by 3-fold when compared to untreated. In addition compound Aloin, an Aloe exudate showed potent Mtb-FtsZ inhibition (IC50 = 16.7 ±â€¯0.4 µM) but exhibited poor anti-mycobacterial activity (>500 µM).


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Proteínas do Citoesqueleto/metabolismo , Mycobacterium smegmatis/efeitos dos fármacos , Alcaloides/farmacologia , Proteínas de Bactérias/genética , Benzodioxóis/farmacologia , Divisão Celular/efeitos dos fármacos , Cumarínicos/farmacologia , Citocinese , Proteínas do Citoesqueleto/genética , Bases de Dados de Compostos Químicos , Avaliação Pré-Clínica de Medicamentos/métodos , GTP Fosfo-Hidrolases/antagonistas & inibidores , Humanos , Simulação de Acoplamento Molecular , Mycobacterium smegmatis/citologia , Mycobacterium tuberculosis/metabolismo , Piperidinas/farmacologia , Alcamidas Poli-Insaturadas/farmacologia , Relação Estrutura-Atividade
19.
Mol Cell ; 74(4): 701-712.e9, 2019 05 16.
Artigo em Inglês | MEDLINE | ID: mdl-30948266

RESUMO

Alternative 3' untranslated regions (3' UTRs) are widespread, but their functional roles are largely unknown. We investigated the function of the long BIRC3 3' UTR, which is upregulated in leukemia. The 3' UTR does not regulate BIRC3 protein localization or abundance but is required for CXCR4-mediated B cell migration. We established an experimental pipeline to study the mechanism of regulation and used mass spectrometry to identify BIRC3 protein interactors. In addition to 3'-UTR-independent interactors involved in known BIRC3 functions, we detected interactors that bind only to BIRC3 protein encoded from the mRNA with the long 3' UTR. They regulate several functions, including CXCR4 trafficking. We further identified RNA-binding proteins differentially bound to the alternative 3' UTRs and found that cooperative binding of Staufen and HuR mediates 3'-UTR-dependent complex formation. We show that the long 3' UTR is required for the formation of specific protein complexes that enable additional functions of BIRC3 protein beyond its 3'-UTR-independent functions.


Assuntos
Proteína 3 com Repetições IAP de Baculovírus/genética , Leucemia/genética , Complexos Multiproteicos/genética , Receptores CXCR4/genética , Regiões 3' não Traduzidas/genética , Linfócitos B/metabolismo , Linfócitos B/patologia , Proteína 3 com Repetições IAP de Baculovírus/química , Movimento Celular/genética , Proteínas do Citoesqueleto/genética , Proteína Semelhante a ELAV 1/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Leucemia/patologia , Complexos Multiproteicos/química , Transporte Proteico , RNA Mensageiro/genética , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética
20.
Gene ; 700: 1-6, 2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-30890479

RESUMO

BACKGROUND AND PURPOSES: Osteoporosis and osteopenia are multifactorial diseases characterized by low bone mineral density (BMD) and are susceptible to genetic and environmental risk factors. The macrophage erythroblast attacher (MAEA) was discovered as a protein to mediate the attachment of erythroid cells to macrophages and is essential for bone marrow hematopoiesis. MAEA is expressed in a wide range of cells and tissues including osteoblasts and osteoclasts. Recent studies have shown that a single nucleotide polymorphism (SNP) rs6815464 (C/G) in the MAEA gene increases the susceptibility of type 2 diabetes mellitus. However, the contribution of MAEA to bone metabolism remains unknown. Therefore, we performed this study to evaluate the association between MAEA polymorphism and low BMD. METHODS: In a cross-sectional study with postmenopausal Japanese women living in the Yokogoshi area, Niigata City, we evaluated whether rs6815464 was associated with low BMD. Blood samples were collected from 353 subjects (age 63.8 ±â€¯5.4 years). The MAEA genotype was determined by TaqMan assay. BMD was assessed by dual-energy X-ray absorptiometry at the lumbar spine (L2-L4), hip and femoral neck. Low BMD was defined as a T-score <-1. RESULTS: The percentage of subjects with low BMD in the lumbar spine, total hip and femoral neck were 71%, 75% and 84% respectively. After adjusting age, BMI, HbA1c, smoking and alcohol consumption, the G-allele carriage was found to be associated with low BMD of total hip (odds ratio = 2.11, 95% CI: 1.14-3.91, P = 0.018), but not of the lumbar spine or femoral neck. CONCLUSION: The MAEA gene polymorphism rs6815464 was associated with low hip BMD in postmenopausal Japanese women.


Assuntos
Grupo com Ancestrais do Continente Asiático/genética , Moléculas de Adesão Celular/genética , Proteínas do Citoesqueleto/genética , Osteoporose Pós-Menopausa/genética , Ossos Pélvicos/diagnóstico por imagem , Polimorfismo de Nucleotídeo Único , Absorciometria de Fóton , Idoso , Densidade Óssea , Estudos Transversais , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Japão , Pessoa de Meia-Idade , Osteoporose Pós-Menopausa/diagnóstico por imagem
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