Scaffold expulsion and genome packaging trigger stabilization of herpes simplex virus capsids.
Herpes simplex virus type 1 (HSV1) capsids undergo extensive structural changes during maturation and DNA packaging. As a result, they become more stable and competent for nuclear egress. To further elucidate this stabilization process, we used biochemical and nanoindentation approaches to analyze the structural and mechanical properties of scaffold-containing (B), empty (A), and DNA-containing (C) nuclear capsids. Atomic force microscopy experiments revealed that A and C capsids were mechanically indistinguishable, indicating that the presence of DNA does not account for changes in mechanical properties during capsid maturation. Despite having the same rigidity, the scaffold-containing B capsids broke at significantly lower forces than A and C capsids. An extraction of pentons with guanidine hydrochloride (GuHCl) increased the flexibility of all capsids. Surprisingly, the breaking forces of the modified A and C capsids dropped to similar values as those of the GuHCl-treated B capsids, indicating that mechanical reinforcement occurs at the vertices. Nonetheless, it also showed that HSV1 capsids possess a remarkable structural integrity that was preserved after removal of pentons. We suggest that HSV1 capsids are stabilized after removal of the scaffold proteins, and that this stabilization is triggered by the packaging of DNA, but independent of the actual presence of DNA.
Roos
Wouter H
WH
Natuur en Sterrenkunde, Vrije Universiteit, De Boelelaan 1081, 1081 HV, Amsterdam, The Netherlands.
Radtke
Kerstin
K
Kniesmeijer
Edward
E
Geertsema
Hylkje
H
Sodeik
Beate
B
Wuite
Gijs J L
GJ
D000818:Animals; D015153:Blotting, Western; D002213:Capsid; D002460:Cell Line; D006224:Cricetinae; D004279:DNA, Viral / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D016679:Genome, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics*; D018625:Microscopy, Atomic Force; D016133:Polymerase Chain Reaction; D019065:Virus Assembly
D004279:DNA, Viral