Structural Studies on Varicella Zoster Virus
Abstract
Little is known about the structure of varicella zoster virus (VZV) except what can be inferred from the more extensively studied herpes simplex virus (HSV). The core of VZV has only been visualized using thin sections of virions or infected cells. By using a unique technique called ion etching, which uses Ar+ to strip away the outer layers of the virus, we have visualized what may be the intact core of the virion. We have seen toroidal structures approximately 70 nm in diameter, similar to those seen in thin section. We have also seen evidence suggesting that the toroids may be open-ended structures instead of closed rings. To identify and further study VZV structural proteins we have cloned the following VZV open reading frames (ORFs) into the vaccinia shuttle vector pSC11: ORF11, ORF33, ORF34, ORF40, ORF47, and ORF66. These ORFs were chosen as potential structural genes based upon knowledge of their HSV homologues. Using these clones we constructed vaccinia recombinants for all but ORF11. Rabbits have been inoculated with these recombinants to create antibodies for use in determining if these are structural proteins, and for studies in cellular localization of the proteins. The recombinant for ORF40, the major capsid protein, will also be used for studies on self assembly of the capsid. Lastly, we have examined the protein kinase (PK) activity which is associated with purified VZ virions. Kinase activity was extracted from the virions and was fractionated using Amicon filters. When added to heat inactivated virions, the activities of the fractions suggested the presence of more than one PK in the virions. Auto phosphorylation studies using purified virions revealed the presence of both cellular and viral PKs. To aid in identifying the viral PK we have cloned VZV genes 47 and 66, which contain eukaryotic PK domains, into the phagemid pBluescript which allows in vitro expression of the genes.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 20, 1990
- Accession Number
- AD1010964
Entities
People
- Jace M. Hougland
Organizations
- Uniformed Services University of the Health Sciences