Earticle

초록

영어

The interferon-induced GTP-binding protein Mx is responsible for a specific antiviral state against a broad spectrum of viral infections that are induced by type-I interferons (IFN α/β) in different vertebrates. In this study, the Mx gene was isolated from the constructed mullet cDNA database. Structural features of mullet Mx (MuMx) were analyzed using different in-silico tools. The pairwise comparison revealed that the MuMx sequence was related to Stegastes partitus Mx with an 83.7% sequence identity, whereas MuMx was clustered into the teleost category in the phylogentic analysis. Sequence alignment showed that the dynamin-type guanine nucleotide- binding domain (G_DYNAMIN_2), central interactive domain (CID), and GTPase effector domain (GED) were conserved among Mx counterparts. The transcriptional expression of MuMx was the highest in blood cells from unchallenged fish. The temporal mRNA profile showed that MuMx expression was significantly elevated in all tissues, including blood, spleen, head kidney, liver, and gills after the injection of polyinosinic-polycytidylic acid (poly I:C) at many time points. Moreover, MuMx expression increased slightly, in the blood, spleen, and head kidney at a few time points after the injection of lipopolysaccharide (LPS) and Lactococcus garvieae (L. garvieae). Results of the subcellular localization analysis confirmed that the MuMx protein was highly expressed in the cytoplasm. The analysis of the gene expression of the viral hemorrhagic septicemia virus (VHSV) under conditions of MuMx overexpression confirmed the significant inhibition of viral transcripts. The cell viability (MTT) assay and VHSV titer quantification with the presence of MuMx indicated a significant reduction in virus replication. Collectively, these findings suggest that Mx is a specific immune-related gene that elicits crucial antiviral functions against viral antigens in the mullet fish.

목차

ABSTRACT
1. Introduction
2. Material and methods
2.1. Construction of mullet complementary DNA (cDNA) library
2.2. Mullet fish rearing and collection of tissues
2.3. Immune challenge experiment
2.4. Extraction of RNA and cDNA synthesis
2.5. Analysis of the expression profile of MuMx
2.6. In silico analysis
2.7. Development of expression vectors
2.8. Maintenance of the FHM cell line, transfection, and virus treatment
2.9. Subcellular localization of MuMx
2.10. Determination of the antiviral activity of MuMx
2.11. Determination of the survival rate of MuMx-overexpressing cells after VHSV infection
2.12. Determination of VHSV titer under the MuMx gene expression
3. Results and discussion
3.1. Structural features of the MuMx sequence
3.2. Homology analysis and the evolutionary relationships of MuMx
3.3. Tissue distribution of MuMx
3.4. Temporal mRNA expression of MuMx
3.5. Assessment of the subcellular localization of MuMx
3.6. Analysis of the antiviral activity of MuMx under conditions of VHSV infection
3.7. Effect of MuMx treatment on the cell survival rate under conditions ofVHSV infection
4. Conclusion
Acknowledgments
References

키워드

저자

• D.M.K.P. Sirisena [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• M.D. Neranjan Tharuka [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• D.S. Liyanage [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• Sumi Jung [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• Myoung-Jin Kim [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• Jehee Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ] Corresponding Author

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]

• 간기

  폐간

• pISSN

  1225-5734

• 수록기간

  1977~2021

• 십진분류

  KDC 454 DDC 551