106-107年度「國立臺灣大學與中央研究院創新性合作計畫」
藉由調控熱休克蛋白MRJ之多聚腺苷酸化機制來發展全方位抗病毒策略
Study of alternative polyadenylation as a regulatory mechanism of mammalian DnaJ (MRJ) and development of a broad-spectrum antiviral strategy through manipulation of MRJ
計畫成員
國立臺灣大學醫學系：黃立民 教授
中央研究院生醫所：譚婉玉 研究員
計畫執行期間：2017, 01, 01-2018, 12, 31
優良研究成果
Emerging and re-emerging viruses continue to threaten humans. The development of timely available antiviral agents and broadspectrum antiviral strategies is important. Therefore, we attempted to identify the strategies to disrupt viral-host interactions or target cellular components required for viral propagation. We have previously reported that the level of the long DNAJB6/MRJ isoform (MRJ-L) in macrophages correlates with the susceptibility of individuals to HIV infection. Therefore, we set out to decipher the molecular mechanism underlying MRJ isoform expression and develop an anti-viral strategy by targeting MRJ isoform. MRJ acts as a protein chaperon and exhibits two splice isoforms that have different roles in human viral infection. In our study, we found that reduction of the polyadenylation factor CstF64 was correlated with the increase of MRJ-L in human macrophage, and then elucidated the mechanism underlying CstF64-modulated MRJ isoform expression. Moreover, we exploited an antisense strategy targeting MRJ-L for virus replication. A morpholino oligonucleotide complementary to the 5’ splice site of MRJ intron 8 downregulated MRJ-L expression and suppressed the replication of not only HIV-1 but also respiratory syncytial virus
(RSV). We demonstrated that downregulation of the MRJ-L level reduced HIV-1 replication as well as the subgenomic mRNA and viral production of RSV. Therefore, our findings that two human health-threatening viruses take advantage of MRJ-L for infection suggest MRJ-L as a potential target for broad-spectrum antiviral strategy (publication 1).
In light of the importance of cellular heat shock proteins including MRJ in viral life cycle and infectivity, we published a review article, in which we discuss how the two MRJ isoforms participate differentially in viral production and virulence, and the possibility for MRJ as a therapeutic target (publication 2).
- Ko, S.-H., Liau, Y.-J., Chi, Y.-H., Lai, M.-J., Chiang, Y.-P., Lu, C.-Y., Chang, L.-Y., Tarn, W.-Y.*, and Huang, L.-M.* (2019) Interference of DNAJB6/MRJ isoform switch by morpholino inhibits replication of HIV-1 and RSV. Mol Therapy-Nucl. Acids 14, 251-261 (co-corresponding with Li-Ming Huang)
- Ko, S.-H., Huang, L.-M.*, and Tarn, W.-Y.* (2019) The host heat shock protein MRJ/DNAJB6 modulates virus infection. Front Microbiol. 10, 2885 (co-corresponding with Li-Ming Huang)