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Search results for: DCTN6

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#31597779   2019/12/12 To Up

Structural Glycoprotein E2 of Classical Swine Fever Virus Interacts with Host Protein Dynactin Subunit 6 (DCTN6) during the Virus Infectious Cycle.

The E2 protein in classical swine fever (CSF) virus (CSFV) is the major virus structural glycoprotein and is an essential component of the viral particle. E2 has been shown to be involved in several functions, including virus adsorption, induction of protective immunity, and virulence in swine. Using the yeast two-hybrid system, we previously identified a swine host protein, dynactin subunit 6 (DCTN6) (a component of the cell dynactin complex), as a specific binding partner for E2. We confirmed the interaction between DCTN6 and E2 proteins in CSFV-infected swine cells by using two additional independent methodologies, i.e., coimmunoprecipitation and proximity ligation assays. E2 residues critical for mediating the protein-protein interaction with DCTN6 were mapped by a reverse yeast two-hybrid approach using a randomly mutated E2 library. A recombinant CSFV mutant, E2ΔDCTN6v, harboring specific substitutions in those critical residues was developed to assess the importance of the E2-DCTN6 protein-protein interaction for virus replication and virulence in swine. CSFV E2ΔDCTN6v showed reduced replication, compared with the parental virus, in an established swine cell line (SK6) and in primary swine macrophage cultures. Remarkably, animals infected with CSFV E2ΔDCTN6v remained clinically normal during the 21-day observation period, which suggests that the ability of CSFV E2 to bind host DCTN6 protein efficiently during infection may play a role in viral virulence. Structural glycoprotein E2 is an important component of CSFV due to its involvement in many virus activities, particularly virus-host interactions. Here, we present the description and characterization of the protein-protein interaction between E2 and the swine host protein DCTN6 during virus infection. The E2 amino acid residues mediating the interaction with DCTN6 were also identified. A recombinant CSFV harboring mutations disrupting the E2-DCTN6 interaction was created. The effect of disrupting the E2-DCTN6 protein-protein interaction was studied using reverse genetics. It was shown that the same amino acid substitutions that abrogated the E2-DCTN6 interaction constituted a critical factor in viral virulence in the natural host, domestic swine. This highlights the potential importance of the E2-DCTN6 protein-protein interaction in CSFV virulence and provides possible mechanisms of virus attenuation for the development of improved CSF vaccines.
M V Borca, E A Vuono, E Ramirez-Medina, P Azzinaro, K A Berggren, M Singer, A Rai, S Pruitt, E B Silva, L Velazquez-Salinas, C Carrillo, D P Gladue

2875 related Products with: Structural Glycoprotein E2 of Classical Swine Fever Virus Interacts with Host Protein Dynactin Subunit 6 (DCTN6) during the Virus Infectious Cycle.

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#31260804   2019/06/28 To Up

Dynactin pathway-related gene expression is altered by aging, but not by vitrification.

The storage of surplus oocytes by cryopreservation (OC) is a widely used tool in assisted reproductive technology, but there is a great debate about the effects of cryopreservation on oocyte competence. It is known that OC may affect meiotic spindles but remains unclear if OC may increase the risk of aneuploidy. The aim of this study was to test the effects of OC and women aging on the expression of cytokinesis-related genes playing an important role in the segregation of chromosomes (DCTN1, DCTN2, DCTN3, DCTN6 and PLK1). Results highlighted that OC do not modify the expression of the selected genes, whereas women aging modulate the expression of all transcripts, confirming that aging is the crucial factor affecting meiosis and aneuploidy risk. A new role for Dynactin and PLK1 was shed in light, providing information on the ageing process in the oocyte which may be associated to reduced fertility.
Marco D'Aurora, Maria Cristina Budani, Sara Franchi, Annalina Sarra, Liborio Stuppia, Gian Mario Tiboni, Valentina Gatta

1588 related Products with: Dynactin pathway-related gene expression is altered by aging, but not by vitrification.

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#29864111   2018/06/04 To Up

Prognostic Value of Dynactin mRNA Expression in Cutaneous Melanoma.

BACKGROUND Dynactin (DCTN) is a multi-subunit protein encoded by DCTN genes for 6 subunits. In different diseases the DCTN genes may have different roles; therefore, we investigated the prognostic potential of DCTN mRNA expression in cutaneous melanoma (CM). MATERIAL AND METHODS Data for DCTN mRNA expression in CM patients were obtained from the OncoLnc database, which contains updated gene expression data for 459 CM patients based on the Cancer Genome Atlas. Kaplan-Meier analysis and a Cox regression model were used to determine overall survival (OS) with calculation of hazard ratios (HRs) and 95% confidence intervals (CIs). RESULTS The multivariate survival analysis showed that individually low expression of DCTN1, DCTN2, and DCTN5 and high expression of DCTN6 were associated with favorable OS (adjusted P=0.008, HR=0.676, 95% CI=0.506-0.903; adjusted P=0.004, HR=0.648, 95% CI=0.485-0.867; adjusted P=0.011, HR=0.686, 95% CI=0.514-0.916; and adjusted P=0.018, HR=0.706, 95% CI=0.530-0.942, respectively). In a joint-effects analysis, combinations of low expression of DCTN1, DCTN2, and DCTN5 and high expression of DCTN6 were found to be more highly correlated with favorable OS (all P<0.05). CONCLUSIONS Our findings suggest that downregulated DCTN1, DCTN2, and DCTN5 and upregulated DCTN6 mRNA expression in CM are associated with favorable prognosis and may represent potential prognostic biomarkers. Moreover, use of the 4 genes in combination can improve the sensitivity for predicting OS in CM patients.
Qiaoqi Wang, Xiangkun Wang, Qian Liang, Shijun Wang, Xiwen Liao, Dong Li, Fuqiang Pan

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#28508063   2017/05/03 To Up

The long noncoding RNA acts as an intranuclear organizing hub for pre-mRNA molecules.

Molecular mechanisms by which long noncoding RNA (lncRNA) molecules may influence cancerous condition are poorly understood. The aberrant expression of lncRNA, encoded within the drosophila gene homolog intron, is correlated with a variety of cancers, including human melanomas. We demonstrate by SHAPE-seq and dChIRP that RNA secondary structure has a core pseudoknotted domain. This lncRNA interacts with the intronic regions of six pre-mRNAs: , , , , , and , all of which have cancer-related functions. Hemizygous knockout of by CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 in melanoma cells significantly decreases lncRNA levels, simultaneously decreases the levels of its interacting pre-mRNA molecules, and decreases anchorage-independent growth rate of cells and the rate of in vivo tumor growth in mouse xenografts. These results provide the first demonstration of an lncRNA's three-dimensional coordinating role in facilitating cancer-related gene expression in human melanomas.
Bongyong Lee, Anupama Sahoo, John Marchica, Erwin Holzhauser, Xiaoli Chen, Jian-Liang Li, Tatsuya Seki, Subramaniam Shyamala Govindarajan, Fatu Badiane Markey, Mona Batish, Sonali J Lokhande, Shaojie Zhang, Animesh Ray, Ranjan J Perera

2969 related Products with: The long noncoding RNA acts as an intranuclear organizing hub for pre-mRNA molecules.

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#20544419   // To Up

Screening and identification of differentially expressed genes from chickens infected with Newcastle disease virus by suppression subtractive hybridization.

Newcastle disease is an important viral infectious disease caused by Newcastle disease virus (NDV), which leads to severe economic losses in the poultry industry worldwide. The molecular mechanisms involved in the pathogenesis of NDV and the host-directed antiviral responses remain poorly understood. In this study, we screened and identified the differentially expressed transcripts from chicken spleen 36 h post NDV infection using suppression subtractive hybridization (SSH). From the SSH library, we obtained 140 significant differentially expressed sequence tags (ESTs), which could be divided into three categories: high homology genes (58), high homology ESTs (62) and novel ESTs (20). The 58 high homology genes could be grouped into nine clusters based on the best known function of their protein products, which involved signalling transduction (HSPC166, PDE7B, GRIA4, GARNL1), transcriptional regulation (ANP32A, LOC423724, SATB1, QKI, ETV6), cellular molecular dynamics (MYLK, MYO7A, DCTN6), cytoskeleton (LAMA4, LAMC1, COL4A1), stress response (DNAJC15, CIRBP), immune response (TIA1, TOX, CMIP), metabolism (RPS15A, RPL32, GLUT8, CYPR21, DPYD, LOC417295), oxidation-reduction (TXN, MSRB3, GCLC), and others. In addition, we found that the 20 novel ESTs provide a clue for the discovery of some new genes associated with infection. In summary, our findings demonstrate previously unrecognized changes in gene transcription that are associated with NDV infection in vivo, and many differentially expressed genes identified in the study clearly merit further investigation. Our data provide new insights into better understanding the molecular mechanism of host-NDV interaction.
Daoliang Lan, Cheng Tang, Mingyi Li, Hua Yue

1359 related Products with: Screening and identification of differentially expressed genes from chickens infected with Newcastle disease virus by suppression subtractive hybridization.

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