The Process of Wrapping Virus Revealed by a Force Tracing Technique and Simulations

Viral entry into the host cell is the first step of virus infection; however, its dynamic process via endocytosis remains largely elusive. Here, the force tracing technique and single particle simulation are combined to investigate the invagination of single human enterovirus 71 (HEV71, a positive single‐stranded RNA virus that is associated with hand, foot, and mouth disease) via cell membranes during its host cell entry. The experimental results reveal that the HEV71 invaginates in membrane vesicles at a force of 58 ± 16 pN, a duration time of 278 ± 68 ms. The simulation further shows that the virus can reach a partially wrapped state very fast, then the upper surface of the virus is covered by the membrane traveling over a long period of time. Combining the experiment with the simulation, the mechanism of membrane wrapping of virus is uncovered, which provides new insights into how the cell is operated to initiate the endocytosis of virus.

[1]  Shixia Wang,et al.  EV71: an emerging infectious disease vaccine target in the Far East? , 2010, Vaccine.

[2]  M. Ho,et al.  Expression, purification and characterization of enterovirus-71 virus-like particles. , 2006, World journal of gastroenterology.

[3]  M. Hallek,et al.  Real-Time Single-Molecule Imaging of the Infection Pathway of an Adeno-Associated Virus , 2001, Science.

[4]  Kuo-Feng Weng,et al.  Viral and host factors that contribute to pathogenicity of enterovirus 71. , 2012, Future microbiology.

[5]  D. Lohr,et al.  Single-molecule recognition imaging microscopy. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Xin Shang,et al.  Size-dependent endocytosis of single gold nanoparticles. , 2011, Chemical communications.

[7]  Yuntao Wu,et al.  Viral exploitation of actin: force-generation and scaffolding functions in viral infection , 2014, Virologica Sinica.

[8]  A. Helenius,et al.  Vaccinia Virus Uses Macropinocytosis and Apoptotic Mimicry to Enter Host Cells , 2008, Science.

[9]  Philip V. Bayly,et al.  Force generation by endocytic actin patches in budding yeast. , 2014, Biophysical journal.

[10]  Huajian Gao,et al.  Mechanics of receptor-mediated endocytosis. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Feng Zhang,et al.  Assembly of endocytic machinery around individual influenza viruses during viral entry , 2004, Nature Structural &Molecular Biology.

[12]  P. McMinn An overview of the evolution of enterovirus 71 and its clinical and public health significance. , 2002, FEMS microbiology reviews.

[13]  H. Netter,et al.  Immunizations with Chimeric Hepatitis B Virus-Like Particles to Induce Potential Anti-Hepatitis C Virus Neutralizing Antibodies , 2007, Antiviral therapy.

[14]  S. Koike,et al.  Scavenger receptor B2 is a cellular receptor for enterovirus 71 , 2009, Nature Medicine.

[15]  Michael J Rust,et al.  Visualizing infection of individual influenza viruses , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[16]  François Nédélec,et al.  Membrane Mechanics of Endocytosis in Cells with Turgor , 2015, PLoS Comput. Biol..

[17]  Benedict Leimkuhler,et al.  Computational Molecular Dynamics: Challenges, Methods, Ideas , 1999, Computational Molecular Dynamics.

[18]  Mingjun Cai,et al.  Recording the dynamic endocytosis of single gold nanoparticles by AFM-based force tracing. , 2015, Nanoscale.

[19]  Jianwei Wang,et al.  Etiology, pathogenesis, antivirals and vaccines of hand, foot, and mouth disease , 2015 .

[20]  C. Y. Liu,et al.  Assembly of human severe acute respiratory syndrome coronavirus-like particles , 2004, Biochemical and Biophysical Research Communications.

[21]  D. Lohr,et al.  Using atomic force microscopy to study chromatin structure and nucleosome remodeling. , 2007, Methods.

[22]  Min Zhang,et al.  Ultrafast Tracking of a Single Live Virion During the Invagination of a Cell Membrane. , 2015, Small.

[23]  B. Dragnea,et al.  Photothermal imaging and measurement of protein shell stoichiometry of single HIV-1 Gag virus-like nanoparticles. , 2011, ACS nano.

[24]  Lucas Pelkmans,et al.  Caveolar endocytosis of simian virus 40 reveals a new two-step vesicular-transport pathway to the ER , 2001, Nature Cell Biology.

[25]  Yuping Shan,et al.  The structure and function of cell membranes examined by atomic force microscopy and single-molecule force spectroscopy. , 2015, Chemical Society reviews.

[26]  Kow-Tong Chen,et al.  An epidemic of enterovirus 71 infection in Taiwan. Taiwan Enterovirus Epidemic Working Group. , 1999, The New England journal of medicine.

[27]  P. Sun,et al.  Formation of virus-like particles from O-type foot-and-mouth disease virus in insect cells using codon-optimized synthetic genes , 2010, Biotechnology Letters.

[28]  T. Ross,et al.  Influenza virus-like particles elicit broader immune responses than whole virion inactivated influenza virus or recombinant hemagglutinin. , 2007, Vaccine.

[29]  D. Makarov,et al.  Theoretical studies of the mechanical unfolding of the muscle protein titin: Bridging the time-scale gap between simulation and experiment , 2003 .

[30]  Huajian Gao,et al.  Physical Principles of Nanoparticle Cellular Endocytosis. , 2015, ACS nano.

[31]  Ferry Kienberger,et al.  Static and Dynamical Properties of Single Poly(Ethylene Glycol) Molecules Investigated by Force Spectroscopy , 2000 .

[32]  Yu-qiang Ma,et al.  Role of physicochemical properties of coating ligands in receptor-mediated endocytosis of nanoparticles. , 2012, Biomaterials.

[33]  Mingjun Cai,et al.  Single-particle tracking of hepatitis B virus-like vesicle entry into cells. , 2011, Small.

[34]  Feng Ruan,et al.  Risk Factors for Hand, Foot, and Mouth Disease and Herpangina and the Preventive Effect of Hand-washing , 2011, Pediatrics.

[35]  Daan Frenkel,et al.  Receptor-mediated endocytosis of nanoparticles of various shapes. , 2011, Nano letters.