Biomedical subjects

Elizabeth E McNamee

Publications and source records attributed to Elizabeth E McNamee.

3 recordsLinked to original sources

PubMed2005

Evidence for a direct interaction between HSV-1 ICP27 and ICP8 proteins.

Herpes simplex virus 1 (HSV-1) ICP27 and ICP8 proteins have both been implicated in the transcription of late genes and regulation of viral gene expression. We showed previously that ICP27 and ICP8 associate with the RNAP II holoenzyme (Zhou and Knipe, J. Virol. 76, 5893-5904). Here, we demonstrate that ICP27 and ICP8 coprecipitate from lysates of HSV-1-infected HEp2 cells and from lysates of insect cells expressing ICP27 and ICP8, the latter being in the absence of other HSV-1 proteins. By expressing and purifying hexahistidine-tagged ICP8 (His-ICP8) and maltose binding protein (MBP)-tagged ICP27 (MBP-27) proteins and performing in vitro immunoprecipitation and pull-down assays, we also demonstrate that ICP27 and ICP8 coprecipitate in the absence of other viral or cellular proteins. Taken together, these data provide evidence that ICP27 and ICP8 interact directly in vitro and in infected cells. We hypothesize that the ICP27-ICP8 interaction plays a role in the stimulation of late gene transcription.

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PubMed2003

Herpes simplex virus replication compartments can form by coalescence of smaller compartments.

Herpes simplex virus (HSV) uses intranuclear compartmentalization to concentrate the viral and cellular factors required for the progression of the viral life cycle. Processes as varied as viral DNA replication, late gene expression, and capsid assembly take place within discrete structures within the nucleus called replication compartments. Replication compartments are hypothesized to mature from a few distinct structures, called prereplicative sites, that form adjacent to cellular nuclear matrix-associated ND10 sites. During productive infection, the HSV single-stranded DNA-binding protein ICP8 localizes to replication compartments. To further the understanding of replication compartment maturation, we have constructed and characterized a recombinant HSV-1 strain that expresses an ICP8 molecule with green fluorescent protein (GFP) fused to its C terminus. In transfected Vero cells that were infected with HSV, the ICP8-GFP protein localized to prereplicative sites in the presence of the viral DNA synthesis inhibitor phosphonoacetic acid (PAA) or to replication compartments in the absence of PAA. A recombinant HSV-1 strain expressing the ICP8-GFP virus replicated in Vero cells, but the yield was increased by 150-fold in an ICP8-complementing cell line. Using the ICP8-GFP protein as a marker for replication compartments, we show here that these structures start as punctate structures early in infection and grow into large, globular structures that eventually fill the nucleus. Large replication compartments were formed by small structures that either moved through the nucleus to merge with adjacent compartments or remained relatively stationary within the nucleus and grew by accretion and fused with neighboring structures.

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PubMed2002

Herpes simplex virus.

Herpes simplex virus (HSV) commonly causes human infections in the orofacial region (HSV-1) and in the genital region (HSV-2). Productive viral infection in mucosal epithelial cells may result in clinical symptoms and is followed by a latent infection within sensory neurons. During productive infection a large number of viral gene products are expressed while during latent infection few or no viral proteins are expressed. Reactivation from latency results in recurrent infections and disease at or near the primary site of infection. Understanding the details of the two stages of the HSV life cycle is a particular focus of current research on HSV. The virus interacts with and modifies numerous host cell functions in both epithelial and neuronal cells, and studies of HSV have enhanced our knowledge of many fundamental processes in eukaryotic cells. Ongoing research continues to uncover novel effects of HSV on cells, and a complete understanding of HSV infection during both productive and latent infection should allow the design of new antiviral agents and vaccines and increased knowledge of basic cell and molecular biology. This review article is designed to provide an introduction to HSV biology and key aspects of the infection cycle.

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