| Item type |
学術雑誌論文 / Journal Article(1) |
| 公開日 |
2019-04-05 |
| タイトル |
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|
タイトル |
Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus |
| 言語 |
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言語 |
eng |
| 資源タイプ |
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資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
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資源タイプ |
journal article |
| ID登録 |
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|
ID登録 |
10.24517/00053821 |
|
ID登録タイプ |
JaLC |
| 著者 |
Kitamura, Kouichi
Que, Lusheng
Shimadu, Miyuki
Koura, Miki
Ishihara, Yuuki
Wakaea, Kousho
Nakamura, Takashi
Watashi, Koichi
Wakita, Takaji
Muramatsu, Masamichi
|
| 著者別表示 |
喜多村, 晃一
島津, 美幸
小浦, 美樹
村松, 正道
|
| 提供者所属 |
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内容記述タイプ |
Other |
|
内容記述 |
金沢大学医薬保健研究域医学系 |
| 書誌情報 |
PLoS Pathogens
en : 14
巻 6,
p. e1007124,
発行日 2018-06-21
|
| ISSN |
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収録物識別子タイプ |
ISSN |
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収録物識別子 |
1553-7366 |
| DOI |
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関連タイプ |
isIdenticalTo |
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識別子タイプ |
DOI |
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関連識別子 |
10.1371/journal.ppat.1007124 |
| 出版者 |
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出版者 |
Public Library of Science |
| 抄録 |
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内容記述タイプ |
Abstract |
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内容記述 |
Hepatitis B virus (HBV) is one of the major etiological pathogens for liver cirrhosis and hepatocellular carcinoma. Chronic HBV infection is a key factor in these severe liver diseases. During infection, HBV forms a nuclear viral episome in the form of covalently closed circular DNA (cccDNA). Current therapies are not able to efficiently eliminate cccDNA from infected hepatocytes. cccDNA is a master template for viral replication that is formed by the conversion of its precursor, relaxed circular DNA (rcDNA). However, the host factors critical for cccDNA formation remain to be determined. Here, we assessed whether one potential host factor, flap structure-specific endonuclease 1 (FEN1), is involved in cleavage of the flap-like structure in rcDNA. In a cell culture HBV model (Hep38.7-Tet), expression and activity of FEN1 were reduced by siRNA, shRNA, CRISPR/Cas9-mediated genome editing, and a FEN1 inhibitor. These reductions in FEN1 expression and activity did not affect nucleocapsid DNA (NC-DNA) production, but did reduce cccDNA levels in Hep38.7-Tet cells. Exogenous overexpression of wild-type FEN1 rescued the reduced cccDNA production in FEN1-depleted Hep38.7-Tet cells. Anti-FEN1 immunoprecipitation revealed the binding of FEN1 to HBV DNA. An in vitro FEN activity assay demonstrated cleavage of 5′-flap from a synthesized HBV DNA substrate. Furthermore, cccDNA was generated in vitro when purified rcDNA was incubated with recombinant FEN1, DNA polymerase, and DNA ligase. Importantly, FEN1 was required for the in vitro cccDNA formation assay. These results demonstrate that FEN1 is involved in HBV cccDNA formation in cell culture system, and that FEN1, DNA polymerase, and ligase activities are sufficient to convert rcDNA into cccDNA in vitro. |
| 権利 |
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|
権利情報 |
Copyright © 2018 Kitamura et al. http://creativecommons.org/licenses/by/4.0/ |
| 著者版フラグ |
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|
出版タイプ |
VoR |
|
出版タイプResource |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| 関連URI |
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識別子タイプ |
URI |
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関連識別子 |
https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1007124 |
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関連名称 |
https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1007124 |
ME-PR-KITAMURA-K-1007124.pdf (3.9 MB)
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