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이숙경, 정유진 우수논문 사례 2017.12.15
작 성 자  : 교학팀 HIT  : 2334
첨부파일  : 2014.8. Jung Y-J HVI miR-BART20-5p targeting Zta and Rta.pdf
MicroRNA miR-BART20-5p Stabilizes Epstein-Barr Virus Latency by Directly Targeting BZLF1 and BRLF1

Yu-Jin Jung, Hoyun Choi, Hyoji Kim, Suk Kyeong Lee
Department of Medical Lifescience, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea

ABSTRACT
Epstein-Barr virus (EBV) is a human herpesvirus associated with various tumors. Rather than going through the lytic cycle, EBV maintains latency by limiting the expression of viral genes in tumors. Viral microRNAs (miRNAs) of some herpesviruses have been reported to directly target immediate early genes and suppress lytic induction. In this study, we investigated whether BamHI-A rightward transcript (BART) miRNAs targeted two EBV immediate early genes, BZLF1 and BRLF1. Bioinformatic analysis predicted that 12 different BART miRNAs would target BRLF1. Of these, the results of a luciferase reporter assay indicated that only one interacted with the 3= untranslated region (UTR) of BRLF1: miR-BART20-5p. miR-BART20-5p’s effect on gene expression involved two putative seed match sites in the BRLF1 3= UTR, but a mutant version of the miRNA, miR-BART20- 5pm, had no effect on expression. As expected from the fact that the entire 3= UTR of BZLF1 resides within the 3= UTR of BRLF1, miR-BART20-5p interacted with the 3= UTR of BZLF1 as well. BZLF1 and BRLF1 mRNA and protein expression was suppressed in cells of an AGS cell line infected with the recombinant Akata strain of EBV (AGS-EBV) transfected with a miR-BART20-5p mimic. The expression of various EBV early proteins was also suppressed by the miR-BART20-5p mimic. In contrast, BZLF1 and BRLF1 expression in AGS-EBV cells transfected with a miR-BART20-5p inhibitor was enhanced. Furthermore, progeny virus
production was suppressed by the miR-BART20-5p mimic and enhanced by the miR-BART20-5p inhibitor in AGS-EBV cells induced for the lytic cycle. Our data suggest that miR-BART20-5p plays a key role in latency maintenance in EBV-associated tumors by directly targeting immediate early genes.
 
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