Project: Research project

Project Details


Herpesvirus are significant human pathogens and cause a variety of disease
syndromes which range in severity from inapparent infection to
life-threatening disease. A hallmark of herpesviruses in their ability to
establish persistent infection. This ability is especially evident in
diseases such as oral or genital herpes simplex infections, in which
periods of latency are punctuated with episodes of active, localized
lesions. Very little is known about the alterations in virus/cell
interactions which result in herpesviral persistent infection rather than
productive, cytocidal infection. As a model for persistent infection,
transformed hamster cell lines have been obtained by infection with equine
herpesvirus type 1 (EHV-1) preparations rich in defective interfering (DI)
particles; small populations (3-20%) of the cells continuously produce both
standard EHV-1 and DI particles. The remaining majority of cells do not
produce viral particles but have been shown to contain portions of the
EHV-1 genome and to express viral antigens. Inoculation of these cell
lines into syngeneic hamsters result in tumor production, and the resulting
tumor cell lines are virus nonproducers. The overall goal of this proposal
is to gain insight into the molecular biology of persistent infection by
comparing the regulation of viral protein synthesis in these persistently
infected cells to that in cytocidal EHV-1 infection and in cells infected
with DI-rich virus. Specifically, the aims are to: 1) further define the
alpha/beta/gamma temporal regulation of viral protein synthesis in EHV-1
cytocidal infection by using metabolic inhibitors such as cyclohexamide,
actinomycin D, Ara C during infections; 2) characterize, through the use of
isotopic labeling, SDS-PAGE, immunoprecipitation and immunoblotting, the
proteins comprising each class with respect to physical properties
(glycosylation, phosphorylation), temporal appearance, stability, cellular
localization, and function; 3) examine DI-rich infections by the same
methods to detect any aberrant regulation (underproduced, overproduced,
novel proteins); 4) characterize viral proteins produced in persistently
infected cells to discern any altered regulation patterns; and 5) identify
and characterize viral proteins expressed in cloned virus non-producer
transformed cells and in tumor cells to gain information as to which viral
gene products may be necessary for transformation versus persistent
StatusNot started