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Kim, Julan,Kim, Ju-Won,Kim, Dong-Gyun,Nam, Bo-Hye,Kim, Young-Ok,Park, Jung Youn,Kong, Hee Jeong Elsevier 2018 FISH AND SHELLFISH IMMUNOLOGY Vol.79 No.-
<P><B>Abstract</B></P> <P>The tripartite motif-containing (TRIM) proteins are involved in a wide range of cellular processes, and the role of TRIM1 in immunity has been explored. However, fundamental studies on fish TRIM1 are lacking. In this study, we cloned and characterized TRIM1 cDNA from the Korean rose bitterling, <I>Rhodeus uyekii</I> (RuTRIM1). Two RuTRIM1 isoforms (RuTRIM1-X1 and RuTRIM1-X2) were identified. The coding sequence (CDS) of RuTRIM1-X1 comprised 2157 bp encoding a 718-aa protein, and the CDS of RuTRIM1-X2 comprised 1929 bp encoding a 642-aa protein. Both RuTRIM1 isoforms contained a RING finger domain, B-box 1, B-box 2, coiled-coil domain, COS box, FN3 motif, and PRY/SPRY domain. The deduced RuTRIM1-X1 and RuTRIM1-X2 proteins had high amino acid identity (76.27–98.89%) with orthologs from various other species, and a phylogenetic tree was constructed. RuTRIM1-X1 and RuTRIM1-X2 mRNA were expressed in all tissues examined, with the highest expression levels detected in the hepatopancreas. During early development, RuTRIM1-X1 and RuTRIM1-X2 mRNA levels changed differently from the gastrula period to the first feeding stage. An <I>in vivo</I> ubiquitination assay showed that RuTRIM1 exhibited RING-dependent E3 ubiquitin ligase activity, mainly by comparing RuTRIM1-X2 to RuTRIM1-X1. The subcellular localization of the two RuTRIM1 protein isoforms was characterized, revealing that they formed aggregates in cytoplasmic bodies in Raw264.7 cells. Interferon-γ/lipopolysaccharide-induced nuclear factor-κB signaling was negatively regulated by RuTRIM1-X1 and RuTRIM1-X2, and the negative effect was reversed in RING deletion mutants. To our knowledge, this is the first study to characterize fish TRIM1, which may play a role in the inflammatory response.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We isolated two TRIM1 isoforms (RuTRIM1-X1 and RuTRIM1-X2) from <I>Rhodeus uyekii.</I> </LI> <LI> RuTRIM1 cDNAs encode polypeptides with the RBCC motif. </LI> <LI> RuTRIM1 proteins share 76.27–98.89% homology with orthologs from other species. </LI> <LI> The subcellular localization and Ub ligase activity of the two RuTRIM1 protein isoforms were characterized. </LI> <LI> RuTRIM1-X1 and RuTRIM1-X2 negatively regulate the IFN-γ/LPS-induced NF-κB signaling. </LI> </UL> </P>
Development and Characterization of a New Cell Line from Olive Flounder Paralichthys olivaceus
Kim, Ju-Won,Oh, Bang Geun,Kim, Julan,Kim, Dong-Gyun,Nam, Bo-Hye,Kim, Young-Ok,Park, Jung Youn,Cheong, JaeHun,Kong, Hee Jeong The Korean Society of Developmental Biology 2018 발생과 생식 Vol.22 No.3
A new embryonic cell line (OFEC-17FEN) derived from olive flounder Paralichthys olivaceus was developed. OFEC-17FEN cells were subcultured for <30 passages over ~200 days. OFEC-17FEN cells had a doubling time of 114.34 h and modal diploid chromosome number was 48. The pluripotency genes POU5f1 and NANOG were expressed in OFEC-17FEN cells. However, the lack of several pluripotency-related genes expression indicates that OFEC-17FEN cells are not stem cells. OFEC-17FEN cells transfected with plasmid pEGFP-c1 exhibited a strong green fluorescent signal at 48 h after transfection. Accordingly, OFEC-17FEN cells may be useful for both basic research and biotechnological application.
Kyung-Hee Kim,Sanghyun Lee,Hyo Sun Jung,Julan Kim,Jong-Won Park,Choul-Ji Park,Hyejin Kim,Woo-Jin Kim,Dain Lee 한국발생생물학회 2020 발생과 생식 Vol.24 No.3
The caspase10 encodes an initiating caspase that plays an important role in the maintaining the cellular homeostasis by regulating the steps involved in the immune response and cell death. We investigated the expression of caspase10 during the different developmental stages and in olive flounder tissues. Caspase10 increased in the late stage of the formation of immune tissue, and high expression was observed in the gills, kidney, skin, and spleen. The current study analyzed the expressional changes of caspase10 in olive flounder infected with viral hemorrhagic septicemia virus (VHSV). One of the major causes of mass mortality, VHSV infection in olive flounder attributes to significant expression of caspase10 in the gills, spleen, skin, and kidneys. The results indicate a close association of caspase10 expression with the immune response to VHSV infection in olive flounder. The observations could form the basis data for exploration of other fish immune system.
Development and Characterization of a New Cell Line from Olive Flounder Paralichthys olivaceus
Ju-Won Kim,Bang Geun Oh,Julan Kim,Dong-Gyun Kim,Bo-Hye Nam,Young-Ok Kim,Jung Youn Park,JaeHun Cheong,Hee Jeong Kong 한국발생생물학회 2018 발생과 생식 Vol.22 No.3
A new embryonic cell line (OFEC-17FEN) derived from olive flounder Paralichthys olivaceus was developed. OFEC-17FEN cells were subcultured for <30 passages over ~200 days. OFEC-17FEN cells had a doubling time of 114.34 h and modal diploid chromosome number was 48. The pluripotency genes POU5f1 and NANOG were expressed in OFEC-17FEN cells. However, the lack of several pluripotency-related genes expression indicates that OFEC-17FEN cells are not stem cells. OFEC-17FEN cells transfected with plasmid pEGFP-c1 exhibited a strong green fluorescent signal at 48 h after transfection. Accordingly, OFEC-17FEN cells may be useful for both basic research and biotechnological application.
Four Members of Heat Shock Protein 70 Family in Korean Rose Bitterling (Rhodeus uyekii)
Jung Hyun Kim,Chun Mae Dong,Julan Kim,Cheul Min An,Hae Ja Baek,Hee Jeong Kong 한국발생생물학회 2015 발생과 생식 Vol.19 No.3
Heat shock protein (HSP) 70, the highly conserved stress protein families, plays important roles in protecting cells against heat and other stresses in most animal species. In the present study, we identified and characterized four Hsp70 (RuHSP4, RuHSC70, RuHSP12A, RuGRP78) family proteins based on the expressed sequence tag (EST) analysis of the Korean rose bitterling R. uyekii cDNA library. The deduced RuHSP70 family has high amino acid identities of 72-99% with those of other species. Phylogenetic analysis revealed that RuHsp70 family clustered with fish groups (HSP4, HSC70, HSP12A, GRP78) proteins. Quantitative RT-PCR analysis showed the specific expression patterns of RuHsp70 family members in the early developmental stages and several tissues in Korean rose bitterling. The expression of 4 groups of Hsp70 family was detected in all tested tissue. Particularly, Hsp70 family of Korean rose bitterling is highly expressed in hepatopancreas and sexual gonad (testis and ovary). The expression of Hsp70 family was differentially regulated in accordance with early development stage of Rhodeus uyekii.
Kong, Hee Jeong,Kim, Julan,Kim, Ju-Won,Kim, Hyun-Chul,Noh, Jae Koo,Kim, Young-Ok,Kim, Woo-Jin,Yeo, Sang-Yeob,Park, Jung Youn The Korean Society of Developmental Biology 2019 발생과 생식 Vol.23 No.1
To develop a promoter capable of driving transgene expression in non-model fish, we identified and characterized the muscle-specific alpha-actin gene in olive flounder, Paralichthys olivaceus (PoACTC1). The regulatory region of PoACTC1 includes putative regulatory elements such as a TATA box, two MyoD binding sites, three CArG boxes, and a CCAAT box. Microinjection experiments demonstrated that the regulatory region of PoACTC1, covering from -2,126 bp to +751 bp, just prior to the start codon, drove the expression of red fluorescent protein in developing zebrafish embryos and hatching olive flounder. These results suggest that the regulatory region of PoACTC1 may be useful in developing a promoter for biotechnological applications such as transgene expression in olive flounder.
Four Members of Heat Shock Protein 70 Family in Korean Rose Bitterling (Rhodeus uyekii)
Kim, Jung Hyun,Dong, Chun Mae,Kim, Julan,An, Cheul Min,Baek, Hae Ja,Kong, Hee Jeong The Korean Society of Developmental Biology 2015 발생과 생식 Vol.19 No.3
Heat shock protein (HSP) 70, the highly conserved stress protein families, plays important roles in protecting cells against heat and other stresses in most animal species. In the present study, we identified and characterized four Hsp70 (RuHSP4, RuHSC70, RuHSP12A, RuGRP78) family proteins based on the expressed sequence tag (EST) analysis of the Korean rose bitterling R. uyekii cDNA library. The deduced RuHSP70 family has high amino acid identities of 72-99% with those of other species. Phylogenetic analysis revealed that RuHsp70 family clustered with fish groups (HSP4, HSC70, HSP12A, GRP78) proteins. Quantitative RT-PCR analysis showed the specific expression patterns of RuHsp70 family members in the early developmental stages and several tissues in Korean rose bitterling. The expression of 4 groups of Hsp70 family was detected in all tested tissue. Particularly, Hsp70 family of Korean rose bitterling is highly expressed in hepatopancreas and sexual gonad (testis and ovary). The expression of Hsp70 family was differentially regulated in accordance with early development stage of Rhodeus uyekii.
Kyung-Hee Kim,Sanghyun Lee,Jong-Won Park,Hyo Sun Jung,Julan Kim,Hyerim Yang,Jeong-Ho Lee,Dain Lee 한국발생생물학회 2021 발생과 생식 Vol.25 No.4
Interferon Regulatory Factor 3 (IRF3) is a member of interferon-regulated transcription factor family and is known to play an important role in the innate immune response against viral infections. In this study, the expression of IRF3 in different tissues, developmental stages, and stocking densities of olive flounder was investigated. The expression of IRF3 was observed to gradually increase in early-stage juvenile fish. The highest expression was observed in later-stage juvenile fish when immune tissues were formed. High IRF3 expression was observed in the muscles and the brain tissues. The expression of IRF3 was observed in fish at different stocking densities after viral hemorrhagic septicemia virus (VHSV) infection. It yielded an interesting expression pattern in the muscles and the brain tissues of fish stocked at low density. These observations can be used as basic data for the study of the expression of immune response-related genes against viruses based on stocking density and immune systems in other fish species.