title: Neuraminidase of Influenza A Virus Binds Lysosome-Associated Membrane Proteins Directly and Induces Lysosome Rupture.
abstract: As a recycling center, lysosomes are filled with numerous acid hydrolase enzymes that break down waste materials and invading pathogens. Recently, lysosomal cell death has been defined as "lysosomal membrane permeabilization and the consequent leakage of lysosome contents into cytosol." Here, we show that the neuraminidase (NA) of H5N1 influenza A virus markedly deglycosylates and degrades lysosome-associated membrane proteins (LAMPs; the most abundant membrane proteins of lysosome), which induces lysosomal rupture, and finally leads to cell death of alveolar epithelial carcinoma A549 cells and human tracheal epithelial cells. The NA inhibitors peramivir and zanamivir could effectively block the deglycosylation of LAMPs, inhibit the virus cell entry, and prevent cell death induced by the H5N1 influenza virus. The NA of seasonal H1N1 virus, however, does not share these characteristics. Our findings not only reveal a novel role of NA in the early stage of the H5N1 influenza virus life cycle but also elucidate the molecular mechanism of lysosomal rupture crucial for influenza virus induced cell death.
  
  
  OBJECTIVE
  The integrity of lysosomes is vital for maintaining cell homeostasis, cellular defense and clearance of invading pathogens. This study shows that the H5N1 influenza virus could induce lysosomal rupture through deglycosylating lysosome-associated membrane proteins (LAMPs) mediated by the neuraminidase activity of NA protein. NA inhibitors such as peramivir and zanamivir could inhibit the deglycosylation of LAMPs and protect lysosomes, which also further interferes with the H5N1 influenza virus infection at early stage of life cycle. This work is significant because it presents new concepts for NA's function, as well as for influenza inhibitors' mechanism of action, and could partially explain the high mortality and high viral load after H5N1 virus infection in human beings and why NA inhibitors have more potent therapeutic effects for lethal avian influenza virus infections at early stage.
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Xiangwu
authors.initials: X
authors.lastname: Ju
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Yiwu
authors.initials: Y
authors.lastname: Yan
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Sheng
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Lifang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Genetic Engineering Laboratory, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Zhu
authors.initials: Z
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Fengming
authors.initials: F
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Kangtai
authors.initials: K
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Yanxu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zou
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Jianchao
authors.initials: J
authors.lastname: Du
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Huandi
authors.initials: H
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Yang
authors.affiliation: Genetic Engineering Laboratory, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.
authors.forename: Huijun
authors.initials: H
authors.lastname: Lu
authors.affiliation: Genetic Engineering Laboratory, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.
authors.forename: Mingyao
authors.initials: M
authors.lastname: Tian
authors.affiliation: Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Dangsheng
authors.initials: D
authors.lastname: Li
authors.affiliation: Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, State Key Laboratory of Medical Molecular Biology, and Department of Immunology, Peking Union Medical College, Tsinghua University, Beijing, China.
authors.forename: Jianming
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Genetic Engineering Laboratory, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China ningyik@126.com jiang@pumc.edu.cn.
authors.forename: Ningyi
authors.initials: N
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, and Department of Biochemistry and Molecular Biology, Peking Union Medical College, Tsinghua University, Beijing, China State Key Laboratory of Biotherapy/Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, Chengdu, China ningyik@126.com jiang@pumc.edu.cn.
authors.forename: Chengyu
authors.initials: C
authors.lastname: Jiang
mesh_terms: D000146:Acids, Carbocyclic; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology*; D003517:Cyclopentanes / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology / Q000821:virology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000821:virology; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D053118:Influenza A Virus, H1N1 Subtype / Q000737:chemistry / Q000201:enzymology; D053124:Influenza A Virus, H5N1 Subtype / Q000737:chemistry / Q000201:enzymology; D051907:Lysosomal Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology* / Q000821:virology; D009439:Neuraminidase / Q000378:metabolism*; D011485:Protein Binding; D059748:Proteolysis; D020545:Respiratory Mucosa / Q000187:drug effects / Q000821:virology; D013045:Species Specificity; D014764:Viral Proteins / Q000378:metabolism*; D053586:Virus Internalization / Q000187:drug effects; D053243:Zanamivir / Q000494:pharmacology
chemical_list: D000146:Acids, Carbocyclic; D003517:Cyclopentanes; D004791:Enzyme Inhibitors; D006146:Guanidines; D051907:Lysosomal Membrane Proteins; D014764:Viral Proteins; D009439:Neuraminidase; D053243:Zanamivir; C414210:peramivir