Charaterization by two-dimensional gel electrophoresis of host proteins whose synthesis is sustained or stimulated during the course of herpes simplex virus type 1 infection

Abstract

Herpes simplex virus type 1 (HSV-1) gene expression is concomitant with a selective shutoff of host protein synthesis. While the synthesis of the vast majority of cellular proteins is inhibited immediately after infection, several cellular proteins continue to be synthesized, even during the late phase of infection. Because these cellular proteins may intervene in the life cycle of the virus, we undertook two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analyses to evaluate the proportion of cellular proteins that is represented by these particular proteins. Human cells were infected with HSV-1. At different times after infection, proteins were labeled with 35S just prior to harvesting. The rate of synthesis of a set of 183 acidic host proteins, as well as that of ribosomal proteins, was measured during the course of infection, after separation by 2-D PAGE. As expected, HSV-1 induces a strong inhibition of host protein synthesis immediately after infection. However, the synthesis of basic ribosomal proteins and that of an unexpected high proportion of the sub-set of cellular proteins analyzed is sustained or stimulated during HSV-1 infection. A 2-D PAGE analysis outlining the expression patterns of these proteins at different times of infection is presented.

Investigating the biology of alpha herpesviruses with MS‐based proteomics

“…Only a small portion of host cell proteins was altered during early stages of infection by either HSV‐1 or PRV. This finding is consistent with the earlier study using only 2DE analysis . Both studies showed that the expression levels of lamin A/C and hnRNP K were affected by infection.…”

“…Systematic studies of viral transcription profiles are useful, but they cannot report on protein composition, stability, or modifications (see for example ). Early studies compared the mock‐infected and HSV‐1‐infected cell proteome using 35 S pulse‐labeling and 2DE . While HSV‐1 infection strongly inhibited host protein synthesis, the synthesis of a number of acidic proteins and basic ribosome proteins was maintained or stimulated.…”

“…Monitoring the global proteomic profile in host cells often obscures subtle alterations in subcellular fractions such as the cytoplasm, ribosomes, and nucleus . Studies utilizing fractionation methods have expanded our knowledge of the host response to α‐HV infection.…”

Proteomic analysis of cells in the early stages of herpes simplex virus type‐1 infection reveals widespread changes in the host cell proteome

“…Although detailed studies have been conducted examining the roles of individual viral proteins during replication, little is known about how these events occur on a global‐proteomic level during infection. Comparative 2‐DE has been used to examine the effect of HSV‐1 infection on host ribosomal proteins 26 and vhs activity 27. Global proteomic studies have been performed on cells transformed by the Epstein–Barr virus (EBV, a member of the gammaherpesvirus family) 28 and more recently on cells infected with Pseudorabies virus (a member of the alphaherpesvirus family) 29.…”

S‐adenosyl methionine decarboxylase activity is required for the outcome of herpes simplex virus type 1 infection and represents a new potential therapeutic target

“…SPD is found in the envelope and SPM in the nucleocapsid (17). We have identified one of the cellular proteins that we have shown previously to escape the virally induced shutoff of cellular protein synthesis (14) as being ornithine decarboxylase (ODC) (unpublished results). ODC is one of the key enzymes of the polyamine biosynthetic pathway.…”