Grass Carp (Ctenopharyngodon idellus) NIMA-Related Kinase 6 Blocks dsRNA-Induced IFN I Response by Targeting IRF3

Accumulating evidence indicates that mammalian NIMA (never in mitosis, gene A)-related kinase 6 (NEK6) plays potential roles during the course of tumorigenesis, but little is known about NEK6 in lower vertebrates. Herein, we reported a mammalian ortholog of NEK6 in grass carp (Ctenopharyngodon idellus) (CiNEK6). Multiple alignment of amino acid sequences and phylogenetic analysis showed that CiNEK6 shares a high level of sequence similarity with its counterparts in birds. CiNEK6 was ubiquitously expressed in all tested tissues, and its expression level was increased under treatment with GCRV (dsRNA virus) or poly I:C (dsRNA analog). Q-PCR and dual-luciferase assays suggested that CiNEK6 overexpression suppressed IFN I activity in CIK cells treated with poly I:C. Knockdown of CiNEK6 resulted in a higher level of IFN I expression in CIK cells treated with poly I:C compared to those which received PBS. Interestingly, analysis of subcellular localization demonstrated that CiNEK6 protein scattered throughout the cytoplasm is gradually congregated together at the edges of karyotheca upon stimulation with poly I:C. Co-IP and co-localization assays suggested that CiNEK6 interacts with CiIRF3 after poly I:C challenge. In poly I:C-treated cells, the phosphorylation of CiIRF3 was increased by CiNEK6 knockdown, but was suppressed by CiNEK6 overexpression, suggesting that CiNEK6 decreases IFN I expression through inhibiting CiIRF3 activity. Cell viability assay, crystal violet staining, and detection of Vp5 also showed that CiNEK6 plays an inhibitory role in IRF3-mediated antiviral responses.

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Comparison of Cantharidin Toxicity in Breast Cancer Cells to Two Common Chemotherapeutics

International Journal of Breast Cancer ◽

10.1155/2014/423059 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 4

Author(s):

Katie M. Kern ◽

Jennifer R. Schroeder

Keyword(s):

Breast Cancer ◽

Cell Viability ◽

Cell Lines ◽

Breast Cancer Cell Lines ◽

Cell Viability Assay ◽

Chemotherapy Drugs ◽

Viability Assay ◽

Systemic Side Effects ◽

High Level ◽

Cancerous Cells

As part of a larger study synthesizing a more directed form of chemotherapy, we have begun to assess the efficacy of different potential toxins that could be delivered locally rather than systemically. In doing so, we hope to reduce the systemic side effects commonly observed, while maintaining a high level of toxicity and eliminating the need for metabolic alterations. In a search for this more efficient method for killing cancerous cells, we have begun studying cantharidin, a toxin used in traditional Chinese medicine, as a potential chemotherapeutic. Using an MTT cell viability assay, the toxicity of cantharidin was compared to both cyclophosphamide and paclitaxel in three different breast cancer cell lines: MCF-7, MDA-MB-231, and SK-BR-3. Increasing the concentration of chemotherapy drugs did decrease cell viability in all cell lines when cantharidin and cyclophosphamide were applied; however differences for paclitaxel were cell-specific. Additionally, cantharidin exhibited the highest decrease in cell viability regardless of cell type, indicating it may be a much more potent and less specific chemotherapeutic. These results will help us move forward in developing a potentially more potent treatment for breast cancer that might eliminate the need for subtype-specific treatments.

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Comprehensive analysis of circRNA expression pattern and circRNA–mRNA–miRNA network in Ctenopharyngodon idellus kidney (CIK) cells after grass carp reovirus (GCRV) infection

Aquaculture ◽

10.1016/j.aquaculture.2019.734349 ◽

2019 ◽

Vol 512 ◽

pp. 734349 ◽

Cited By ~ 2

Author(s):

Bo Liu ◽

Rui Yuan ◽

Zi Liang ◽

Tingting Zhang ◽

Min Zhu ◽

...

Keyword(s):

Expression Pattern ◽

Grass Carp ◽

Comprehensive Analysis ◽

Ctenopharyngodon Idellus ◽

Grass Carp Reovirus ◽

Cik Cells

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Identification and genomic characterization of a novel fish reovirus, Hubei grass carp disease reovirus, isolated in 2009 in China

Journal of General Virology ◽

10.1099/vir.0.054767-0 ◽

2013 ◽

Vol 94 (10) ◽

pp. 2266-2277 ◽

Cited By ~ 47

Author(s):

Yuding Fan ◽

Shujing Rao ◽

Lingbing Zeng ◽

Jie Ma ◽

Yong Zhou ◽

...

Keyword(s):

Amino Acid ◽

Grass Carp ◽

Sequence Similarity ◽

Amino Acid Level ◽

Amino Acid Sequences ◽

Full Genome Sequence ◽

Total Size ◽

The Family ◽

Genomic Characterization

A novel fish reovirus, Hubei grass carp disease reovirus (HGDRV; formerly grass carp reovirus strain 104, GCRV104), was isolated from diseased grass carp in China in 2009 and the full genome sequence was determined. This reovirus was propagated in a grass carp kidney cell line with a typical cytopathic effect. The total size of the genome was 23 706 bp with a 51 mol% G+C content, and the 11 dsRNA segments encoded 12 proteins (two proteins encoded by segment 11). A nucleotide sequence similarity search using blastn found no significant matches except for segment 2, which partially matched that of the RNA-dependent RNA polymerase (RdRp) from several viruses in the genera Aquareovirus and Orthoreovirus of the family Reoviridae. At the amino acid level, seven segments (Seg-1 to Seg-6, and Seg-8) matched with species in the genera Aquareovirus (15–46 % identities) and Orthoreovirus (12–44 % identities), while for four segments (Seg-7, Seg-9, Seg-10 and Seg-11) no similarities in these genera were found. Conserved terminal sequences, 5′-GAAUU----UCAUC-3′, were found in each HGDRV segment at the 5′ and 3′ ends, and the 5′-terminal nucleotides were different from any known species in the genus Aquareovirus. Phylogenetic analysis based on RdRp amino acid sequences from members of the family Reoviridae showed that HGDRV clustered with aquareoviruses prior to joining a branch common with orthoreoviruses. Based on these observations, we propose that HGDRV is a new species in the genus Aquareovirus that is distantly related to any known species within this genus.

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Cloning and characterization of two glutathione S-transferases from a DDT-resistant strain of Anopheles gambiae

Biochemical Journal ◽

10.1042/bj3240097 ◽

1997 ◽

Vol 324 (1) ◽

pp. 97-102 ◽

Cited By ~ 94

Author(s):

Hilary RANSON ◽

La-aied PRAPANTHADARA ◽

Janet HEMINGWAY

Keyword(s):

Anopheles Gambiae ◽

Resistant Strain ◽

Sequence Similarity ◽

Amino Acid Sequences ◽

Class I ◽

Kinetic Properties ◽

Coding Region ◽

High Sequence Similarity ◽

Glutathione S Transferases ◽

High Level

Two cDNA species, aggst1-5 and aggst1-6, comprising the entire coding region of two distinct glutathione S-transferases (GSTs) have been isolated from a 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (DDT) resistant strain (ZANDS) of Anopheles gambiae. The nucleotide sequences of these cDNA species share 80.2% identity and their derived amino acid sequences are 82.3% similar. They have been classified as insect class I GSTs on the basis of their high sequence similarity to class I GSTs from Drosophila melanogaster and Musca domestica and they are localized to a region of an An. gambiae chromosome known to contain further class I GSTs. The genes aggst1-5 and aggst1-6 were expressed at high levels in Escherichia coli and the recombinant GSTs were purified by affinity chromatography and characterized. Both agGST1-5 and agGST1-6 showed high activity with the substrates 1-chloro-2,4-dinitrobenzene and 1,2-dichloro-4-nitrobenzene but negligible activity with the mammalian theta class substrates, 1,2-epoxy-3-(4-nitrophenoxy)propane and p-nitrophenyl bromide. Despite their high level of sequence identity, agGST1-5 and agGST1-6 displayed different kinetic properties. Both enzymes were able to metabolize DDT and were localized to a subset of GSTs that, from earlier biochemical studies, are known to be involved in insecticide resistance in An. gambiae. This subset of enzymes is one of three in which the DDT metabolism levels are elevated in resistant insects.

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Molecular characterization of ER Stress sensor IRE1α and IRE1β and their expression analysis in response to different fatty acid in grass carp (Ctenopharyngodon idellus)

10.21203/rs.3.rs-167447/v1 ◽

2021 ◽

Author(s):

Zhiguang Chang ◽

Minghui Yang ◽

Hong Ji

Keyword(s):

Fatty Acids ◽

Fatty Acid ◽

Er Stress ◽

Grass Carp ◽

Transmembrane Helix ◽

Specific Inhibitor ◽

Ctenopharyngodon Idellus ◽

Cik Cells ◽

Physiological Processes ◽

Evolutionarily Conserved

Abstract Inositol-requiring enzyme 1 (IRE1) is the most evolutionarily conserved endoplasmic reticulum (ER) membrane protein. Here, we identified and analyzed two IRE isoforms (IRE1α and IRE1β) in grass carp ( Ctenopharyngodon idellus ). The coding sequences of IRE1α and IRE1β were 3096 and 2835 nucleotides, and they encoded proteins of 1031 and 944 amino acids, respectively. Each of the two IRE1s proteins had four PPQ domains, a transmembrane helix region, an S_TKc and a PUG domain, which were relatively conserved in comparison to mammals. qRT-PCR revealed that IRE1α was the highest in liver and brain, whereas the highest expression of IRE1β expressed was found in the hindgut, which may be closely related with their role. Moreover, we have compared the effect of saturated (palmitic acid, PA) , monounsaturated fatty acid (oleic acid, OA) and polyunsaturated fatty acids (Docosahexaenoic acid, DHA) on ER stress in CIK cells, and found that PA and DHA induced UPR-related gene expression in dose-dependent , and OA increased expression of IRE1α and decreased expression of IRE1β, ATF4 and ATF6 at lower doses. Additionally, our studies revealed that blocking IRE1α pathway using specific inhibitor 4μ8c, subsequently reversed PA-induced autophagy and inflammation, indicating the role of IRE1α in mediating PA-induced CIK cells. Taken together, the study demonstrated the IRE1α and IRE1β that likely have an important role in physiological processes induced by fatty acids in grass carp.

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Lycopene alleviates sulfamethoxazole-induced hepatotoxicity in grass carp (Ctenopharyngodon idellus) via suppression of oxidative stress, inflammation and apoptosis

Food & Function ◽

10.1039/d0fo01638a ◽

2020 ◽

Vol 11 (10) ◽

pp. 8547-8559

Author(s):

Hongjing Zhao ◽

Yu Wang ◽

Mengyao Mu ◽

Menghao Guo ◽

Hongxian Yu ◽

...

Keyword(s):

Oxidative Stress ◽

Secondary Metabolites ◽

Human Health ◽

Grass Carp ◽

Aquatic Environment ◽

Ctenopharyngodon Idellus

Antibiotics are used worldwide to treat diseases in humans and other animals; most of them and their secondary metabolites are discharged into the aquatic environment, posing a serious threat to human health.

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