表皮生长因子受体是引发乙型肝炎病毒内化的宿主进入辅助

StephenW

Proc Natl Acad Sci U S A. 2019 Apr 5. pii: 201811064. doi: 10.1073/pnas.1811064116. [Epub ahead of print]
Epidermal growth factor receptor is a host-entry cofactor triggering hepatitis B virus internalization.
Iwamoto M1,2, Saso W1,3, Sugiyama R1, Ishii K4, Ohki M5, Nagamori S6, Suzuki R1, Aizaki H1, Ryo A7, Yun JH8, Park SY5, Ohtani N9, Muramatsu M1, Iwami S2,10,11, Tanaka Y12, Sureau C13, Wakita T1, Watashi K14,10,11,15.
Author information

1
    Department of Virology II, National Institute of Infectious Diseases, 162-8640 Tokyo, Japan.
2
    Mathematical Biology Laboratory, Department of Biology, Faculty of Sciences, Kyushu University, 812-8581 Fukuoka, Japan.
3
    The Institute of Medical Science, The University of Tokyo, 108-8639 Tokyo, Japan.
4
    Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, 162-8640 Tokyo, Japan.
5
    Drug Design Laboratory, Yokohama City University Graduate School of Medical Life Science, 230-0045 Yokohama, Japan.
6
    Department of Collaborative Research, Nara Medical University, 634-8521 Kashihara, Japan.
7
    Department of Microbiology, Yokohama City University School of Medicine, 236-0004 Yokohama, Japan.
8
    Department of Biochemistry, College of Life Science & Biotechnology, Yonsei University, 03722 Seoul, South Korea.
9
    Graduate School of Medicine, Osaka City University, 545-8585 Osaka, Japan.
10
    Core Research for Evolutional Science and Technology (CORE), Japan Science and Technology Agency (JST), 332-0012 Saitama, Japan.
11
    MIRAI, Japan Science and Technology, 332-0012 Saitama, Japan.
12
    Department of Virology and Liver Unit, Nagoya City University Graduate School of Medicinal Sciences, 467-8601 Nagoya, Japan.
13
    Laboratoire de Virologie Moléculaire, Institut National de la Transfusion Sanguine, 75739 Paris, France.
14
    Department of Virology II, National Institute of Infectious Diseases, 162-8640 Tokyo, Japan; [email protected].
15
    Department of Applied Biological Science, Tokyo University of Science, 278-8510 Noda, Japan.

Abstract

Sodium taurocholate cotransporting polypeptide (NTCP) is a host cell receptor required for hepatitis B virus (HBV) entry. However, the susceptibility of NTCP-expressing cells to HBV is diverse depending on the culture condition. Stimulation with epidermal growth factor (EGF) was found to potentiate cell susceptibility to HBV infection. Here, we show that EGF receptor (EGFR) plays a critical role in HBV virion internalization. In EGFR-knockdown cells, HBV or its preS1-specific fluorescence peptide attached to the cell surface, but its internalization was attenuated. PreS1 internalization and HBV infection could be rescued by complementation with functional EGFR. Interestingly, the HBV/preS1-NTCP complex at the cell surface was internalized concomitant with the endocytotic relocalization of EGFR. Molecular interaction between NTCP and EGFR was documented by immunoprecipitation assay. Upon dissociation from functional EGFR, NTCP no longer functioned to support viral infection, as demonstrated by either (i) the introduction of NTCP point mutation that disrupted its interaction with EGFR, (ii) the detrimental effect of decoy peptide interrupting the NTCP-EGFR interaction, or (iii) the pharmacological inactivation of EGFR. Together, these data support the crucial role of EGFR in mediating HBV-NTCP internalization into susceptible cells. EGFR thus provides a yet unidentified missing link from the cell-surface HBV-NTCP attachment to the viral invasion beyond the host cell membrane.
KEYWORDS:

EGFR; HBV; NTCP; entry; transporter

PMID:
    30952782
DOI:
    10.1073/pnas.1811064116

StephenW

Proc Natl Acad Sci U S A. 2019 Apr 5. pii：201811064.doi：10.1073 / pnas.1811064116。 [印刷前的电子版]
表皮生长因子受体是引发乙型肝炎病毒内化的宿主进入辅助因子。
Iwamoto M1,2，Saso W1,3，Sugiyama R1，Ishii K4，Ohki M5，Nagamori S6，Suzuki R1，Aizaki H1，Ryo A7，Yun JH8，Park SY5，Ohtani N9，Muramatsu M1，Iwami S2,10,11，田中Y12，Sureau C13，Wakita T1，Watashi K14,10,11,15。
作者信息

1
    国立传染病研究所病毒学系II，162-8640，日本东京。
2
    九州大学理学部生物系数学生物学实验室，812-8581，日本福冈。
3
    东京大学医学研究所，108-8639，日本东京。
4
    国立传染病研究所质量保证和放射防护系，162-8640，日本东京。
五
    横滨市立大学医学生命科学研究科药物设计实验室，230-0045横滨，日本。
6
    奈良医科大学合作研究系，634-8521 Kashihara，日本。
7
    横滨市立大学医学部微生物学系，236-0004横滨，日本。
8
    延世大学生命科学与生物技术学院生物化学系，03722韩国首尔
9
    大阪市立大学医学研究科，545-8585大阪，日本。
10
    核心研究进化科学与技术（CORE），日本科学技术厅（JST），332-0012埼玉县，日本。
11
    MIRAI，日本科学技术，332-0012埼玉县，日本。
12
    名古屋市立大学医学研究科病毒学与肝科，日本名古屋市467-8601。
13
    Laboratoire deVirologieMoléculaire，Institut National de la Transfusion Sanguine，75739 Paris，France。
14
    国立传染病研究所病毒学系II，日本东京162-8640; [email protected]。
15
    东京理科大学应用生物科学系，278-8510 Noda，Japan。

抽象

牛磺胆酸钠协同转运多肽（NTCP）是乙型肝炎病毒（HBV）进入所需的宿主细胞受体。然而，NTCP表达细胞对HBV的易感性根据培养条件而不同。发现用表皮生长因子（EGF）刺激可增强细胞对HBV感染的易感性。在这里，我们表明EGF受体（EGFR）在HBV病毒粒子内化中起关键作用。在EGFR敲低细胞中，HBV或其preS1特异性荧光肽附着于细胞表面，但其内化作用减弱。通过功能性EGFR的互补可以挽救PreS1内化和HBV感染。有趣的是，细胞表面的HBV / preS1-NTCP复合物被内化，伴随着EGFR的内吞重定位。通过免疫沉淀测定记录NTCP和EGFR之间的分子相互作用。在功能性EGFR解离后，NTCP不再起到支持病毒感染的作用，如（i）引入NTCP点突变破坏其与EGFR的相互作用所证明的，（ii）诱饵肽中断NTCP-EGFR相互作用的不利影响，或（iii）EGFR的药理学灭活。总之，这些数据支持EGFR在介导HBV-NTCP内化到易感细胞中的关键作用。因此，EGFR提供了从细胞表面HBV-NTCP附着到宿主细胞膜之外的病毒入侵的尚未鉴定的缺失环节。
关键词：

EGFR; HBV; NTCP;条目;转运

结论：
    30952782
DOI：
    10.1073 / pnas.1811064116