Herpes simplex virus immediate early proteins and effects on translation

Herpes simplex virus (HSV) gene regulation requires a complex network of viral and cellular protein interactions. HSV expresses its proteins in a temporal cascade in which immediate early (IE) proteins are required for the expression of early (E) proteins, and both IE and E, and viral DNA replication are required for the expression of late (L) proteins. The IE proteins of HSV are mainly involved in regulating viral gene expression, and this regulation is carried out in multiple steps. IE proteins ICP27 and ICP22 were initially shown to regulate gene expression at the level of transcription, however continued studies have shown their involvement in additional aspects of gene regulation. ICP27 is required for efficient viral DNA replication, and the expression of a subset of viral E and L genes. ICP27 associates with a variety of host cell proteins, and many functions of ICP27 are mediated through these interactions. In this dissertation I focused on one aspect of ICP27 gene regulation, translation. We used a proteomics approach to identify novel associations with cellular translation factors elF3 and PABP. We examined protein synthesis rates in comparison to mRNA accumulation, and determined that ICP27 increases translation of a subset of viral L mRNAs. This function requires ICP27 to have an intact C-terminus. We next examined localization of PABP during infection, and determined that PABP localizes to nuclear SC35 domains at the periphery of viral replication compartments. Our initial assumption was that ICP27 was required for re-localization of PABP, because ICP27 associated with PABP in our proteomics studies. However, further examination determined that IE HSV protein ICP22 into SC35 domains induces re-localization of PABP. As the study progressed it became clear that re-localization of PABP paralleled reported ICP22-induced modifications to RNAP II thought to promote inhibition of cellular transcription, as well as facilitate an alternative mechanism of viral transcription. In this dissertation I present data demonstrating that the re-localization of PABP during HSV infection correlates with the loss of ser2P RNAP II induced by ICP22. This dissertation provides further information towards defining a role for ICP27 and ICP22 in regulating viral gene expression.