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Molecular cloning and characterization of Aos1 and Uba2 from the orange-spotted grouper (Epinephelus coioides)
Wei, Jingguang; Li, Chen; Zhang, Xin; Zhou, Sheng; Zang, Shaoqing1,2; Wei, Shina; Qin, Qiwei3
2018
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume81Pages:343
AbstractSmall ubiquitin-related modifiers (SUMOs) are post-translationally conjugated to other proteins and are essential regulators of a wide range of cellular processes. Covalent attachment of SUMO requires an enzymatic cascade consisting of a single E1-activating enzyme (Aos1 and Uba2 heterodimer), a single E2-conjugating enzyme (Ubc9), and one of several E3 ligases that facilitate transfer of SUMO from Ubc9 to the substrate. In the present study, the Aos1 and Uba2 homologues (EcAos1 and EcUba2) from the orange-spotted grouper (Epinephelus coioides) were cloned and their possible roles in fish immunity were analyzed. The open reading frame (ORF) of EcAos1 contains 1050 base pairs (bp) encoding a 350 amino acid protein with a predicted molecular mass of 38.97 kDa EcAos1 has a nuclear localization signal (NLS) at residues 193-203. The ORF of EcUba2 contains 1950 bp encoding a 650 amino acid protein with a predicted molecular mass of 71.3 kDa EcUba2 has a NLS at residues 608-630. Quantitative real-time polymerase chain reaction analysis indicated that both EcAos1 and EcUba2 were distributed in all examined tissues. The expression levels of EcAos1 and EcUba2 in the spleen and head kidney of E. coioides were differentially up-regulated when challenged with polyinosine-polycytidylic acid. Green fluorescence of both pEGFP-C1-EcAos1 and pEGFP-C1-EcUba2 was distributed in the nucleus of GS cells. When the NLSs of EcAos1 and EcUba2 were deleted, the cellular localizations all changed. Over-expression of EcAos1 and EcUba2 inhibited red-spotted grouper nervous necrosis virus infection and replication. These results are important for better understanding of the SUMO pathway in fish and provide insights into the regulatory mechanism of viral infection in E. coioides under farmed conditions.
DepartmentLMB
KeywordEpinephelus coioides Aos1 Uba2 RGNNV Cellular localization
Document Type期刊论文
Identifierhttp://ir.scsio.ac.cn/handle/344004/17631
Collection中科院海洋生物资源可持续利用重点实验室
Affiliation1.South China Agr Univ, Coll Marine Sci, Guangzhou 510642, Guangdong, Peoples R China
2.Chinese Acad Sci, South China Sea Inst Oceanol, CAS Key Lab Trop Marine Bioresources & Ecol, Guangzhou 510301, Guangdong, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266000, Peoples R China
Recommended Citation
GB/T 7714
Wei, Jingguang,Li, Chen,Zhang, Xin,et al. Molecular cloning and characterization of Aos1 and Uba2 from the orange-spotted grouper (Epinephelus coioides)[J]. FISH & SHELLFISH IMMUNOLOGY,2018,81:343, 353.
APA Wei, Jingguang.,Li, Chen.,Zhang, Xin.,Zhou, Sheng.,Zang, Shaoqing.,...&Qin, Qiwei.(2018).Molecular cloning and characterization of Aos1 and Uba2 from the orange-spotted grouper (Epinephelus coioides).FISH & SHELLFISH IMMUNOLOGY,81,343.
MLA Wei, Jingguang,et al."Molecular cloning and characterization of Aos1 and Uba2 from the orange-spotted grouper (Epinephelus coioides)".FISH & SHELLFISH IMMUNOLOGY 81(2018):343.
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