Mechanical disassembly of single virus particles reveals kinetic intermediates predicted by theory.
New experimental approaches are required to detect the elusive transient intermediates predicted by simulations of virus assembly or disassembly. Here, an atomic force microscope (AFM) was used to mechanically induce partial disassembly of single icosahedral T=1 capsids and virions of the minute virus of mice. The kinetic intermediates formed were imaged by AFM. The results revealed that induced disassembly of single minute-virus-of-mice particles is frequently initiated by loss of one of the 20 equivalent capsomers (trimers of capsid protein subunits) leading to a stable, nearly complete particle that does not readily lose further capsomers. With lower frequency, a fairly stable, three-fourths-complete capsid lacking one pentamer of capsomers and a free, stable pentamer were obtained. The intermediates most frequently identified (capsids missing one capsomer, capsids missing one pentamer of capsomers, and free pentamers of capsomers) had been predicted in theoretical studies of reversible capsid assembly based on thermodynamic-kinetic models, molecular dynamics, or oligomerization energies. We conclude that mechanical manipulation and imaging of simple virus particles by AFM can be used to experimentally identify kinetic intermediates predicted by simulations of assembly or disassembly.
Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid), Departamento de Física de la Materia Condensada C-III, Universidad Autónoma de Madrid, Madrid, Spain.
Milagros
M
Castellanos
Rebeca
R
Pérez
Pablo J P
PJ
Carrillo
Pedro J
PJ
de Pablo
Mauricio G
MG
Mateu
D000818:Animals; D002213:Capsid / Q000378:metabolism; D002460:Cell Line; D007700:Kinetics; D051379:Mice; D018625:Microscopy, Atomic Force; D008915:Minute Virus of Mice / Q000737:chemistry / Q000502:physiology; D008954:Models, Biological; D008958:Models, Molecular; D013314:Stress, Mechanical; D013816:Thermodynamics; D014771:Virion / Q000502:physiology*; D019065:Virus Assembly / Q000502:physiology*