title: Disulfide bond formation contributes to herpes simplex virus capsid stability and retention of pentons.
abstract: Disulfide bonds reportedly stabilize the capsids of several viruses, including papillomavirus, polyomavirus, and simian virus 40, and have been detected in herpes simplex virus (HSV) capsids. In this study, we show that in mature HSV-1 virions, capsid proteins VP5, VP23, VP19C, UL17, and UL25 participate in covalent cross-links, and that these are susceptible to dithiothreitol (DTT). In addition, several tegument proteins were found in high-molecular-weight complexes, including VP22, UL36, and UL37. Cross-linked capsid complexes can be detected in virions isolated in the presence and absence of N-ethylmaleimide (NEM), a chemical that reacts irreversibly with free cysteines to block disulfide formation. Intracellular capsids isolated in the absence of NEM contain disulfide cross-linked species; however, intracellular capsids isolated from cells pretreated with NEM did not. Thus, the free cysteines in intracellular capsids appear to be positioned such that disulfide bond formation can occur readily if they are exposed to an oxidizing environment. These results indicate that disulfide cross-links are normally present in extracellular virions but not in intracellular capsids. Interestingly, intracellular capsids isolated in the presence of NEM are unstable; B and C capsids are converted to a novel form that resembles A capsids, indicating that scaffold and DNA are lost. Furthermore, these capsids also have lost pentons and peripentonal triplexes as visualized by cryoelectron microscopy. These data indicate that capsid stability, and especially the retention of pentons, is regulated by the formation of disulfide bonds in the capsid.
authors.affiliation: Departments of Molecular, Microbial, and Structural Biology, The University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030, USA.
authors.forename: Renata
authors.initials: R
authors.lastname: Szczepaniak
authors.forename: Jacob
authors.initials: J
authors.lastname: Nellissery
authors.forename: Joshua A
authors.initials: JA
authors.lastname: Jadwin
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Makhov
authors.forename: Athena
authors.initials: A
authors.lastname: Kosinski
authors.forename: James F
authors.initials: JF
authors.lastname: Conway
authors.forename: Sandra K
authors.initials: SK
authors.lastname: Weller
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry* / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004220:Disulfides / Q000378:metabolism*; D004229:Dithiothreitol / Q000378:metabolism; D005033:Ethylmaleimide / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D008958:Models, Molecular; D019163:Reducing Agents / Q000378:metabolism; D014709:Vero Cells; D014771:Virion / Q000648:ultrastructure
chemical_list: D036022:Capsid Proteins; D004220:Disulfides; D019163:Reducing Agents; D005033:Ethylmaleimide; D004229:Dithiothreitol