Biomedical subjects

Christine M Sanfilippo

Publications and source records attributed to Christine M Sanfilippo.

4 recordsLinked to original sources

PubMed2006

ICP0 gene expression is a herpes simplex virus type 1 apoptotic trigger.

Apoptosis is a highly regulated programmed cell death process which is activated during normal development and by various stimuli, such as viral infection, which disturb cellular metabolism and physiology. That herpes simplex virus type 1 (HSV-1) induces apoptosis but then prevents its killing of infected cells is well-established. However, little is known about the viral factor/event which triggers the apoptotic process. We previously reported that infections with either (i) a temperature-sensitive virus at its nonpermissive temperature which does not inject viral DNA into nuclei or (ii) various UV-inactivated wild-type viruses do not result in the induction of apoptosis (C. M. Sanfilippo, F. N. W. Chirimuuta, and J. A. Blaho, J. Virol. 78:224-239, 2004). This indicates that virus receptor binding/attachment to cells, membrane fusion, virion disassembly/tegument dispersal, virion RNAs, and capsid translocation to nuclei are not responsible for induction and implicates viral immediate-early (IE) gene expression in the process. Here, we systematically evaluated the contribution of each IE gene to the stimulation of apoptosis. Using a series of viruses individually deleted for alpha27, alpha4, and alpha22, we determined that these genes are not required for apoptosis induction but rather that their products play roles in its prevention, likely through regulatory effects. Sole expression of alpha0 acted as an "apoptoxin" that was necessary and sufficient to trigger the cell death cascade. Importantly, results using a recombinant virus which contains a stop codon in alpha0 showed that it was not the ICP0 protein which acted as the apoptotic inducer. Based on these findings, we propose that alpha0 gene expression acts as an initial inducer of apoptosis during HSV-1 infection. This represents the first description of apoptosis induction in infected cells triggered as a result of expression of a single viral gene. Expression of apoptotic viral genes is a unique mechanism through which human pathogens may modulate interactions with their host cells.

Active Transport, Cell Nucleus↗

PubMed2004

Herpes simplex virus 1 infection is required to produce ICP27 immunoreactive triplet forms when ribosomal aminoacyl-tRNA translocation is blocked by cycloheximide.

Infected cell protein (ICP) 27 is an essential herpes simplex virus type 1 (HSV-1) phosphoprotein required for optimal viral DNA and early or late gene synthesis. Three slow-migrating immunoreactive species were detected using multiple anti-ICP27 antibodies following HSV-1 infection of HEp-2 and Vero cells in the presence of cycloheximide (CHX). Generation of the protein triplet moieties required transcription of the alpha27 gene. These forms were observed following infection with a series of recombinant viruses that produce truncated ICP27 polypeptides, suggesting that alternative splicing is not involved in the process. These ICP27 species were not observed following translation inhibition by puromycin (PUR). Synthesis of the triplet occurred by 6 hpi and CHX addition as late as 3 hpi still enabled their production. That the ICP27 species were detected in uninfected ICP27-expressing cells without CHX, but not in its presence, suggests a mechanism in which virus infection is required to produce the forms when ribosomal aminoacyl-transfer RNA (tRNA) translocation is blocked.

Animals↗

PubMed2004

Herpes simplex virus type 1 immediate-early gene expression is required for the induction of apoptosis in human epithelial HEp-2 cells.

Wild-type herpes simplex virus type 1 (HSV-1) induces apoptosis in human epithelial HEp-2 cells, but infected cell proteins produced later in infection block the process from killing the cells. Thus, HSV-1 infection in the presence of the translational inhibitor cycloheximide (CHX) results in apoptosis. Our specific goal was to gain insight as to the viral feature(s) responsible for triggering apoptosis during HSV-1 infection. We now report the following. (i) No viral protein synthesis or death factor processing was detected after infection with HSV-1(HFEMtsB7) at 39.5 degrees C; this mutant virus does not inject its virion DNA into the nucleus at this nonpermissive temperature. (ii) No death factor processing or apoptotic morphological changes were detected following infection with UV-irradiated, replication-defective viruses possessing transcriptionally active incoming VP16. (iii) Addition of the transcriptional inhibitor actinomycin D prevented death factor processing upon infection with the apoptotic, ICP27-deletion virus HSV-1(vBSDelta27). (iv) Apoptotic morphologies and death factor processing were not observed following infection with HSV-1(d109), a green fluorescent protein-expressing recombinant virus possessing deletions of all five immediate-early (IE) (or alpha) genes. (v) Finally, complete death factor processing was observed upon infection with the VP16 transactivation domain-mutant HSV-1(V422) in the presence of CHX. Based on these findings, we conclude that (vi) the expression of HSV-1 alpha/IE genes is required for the viral induction of apoptosis and (vii) the transactivation activity of VP16 is not necessary for this induction.

Animals↗

PubMedDate not supplied

The facts of death.

Cell death is a physiological process critical for organismal development and required for the removal of damaged cells. Apoptosis, the purposeful killing of cells, is highly regulated, and abnormalities in apoptotic control mechanisms contribute to an assortment of human diseases. Depending on the nature of the death signal, separate independent signaling pathways become activated that coordinately participate in the execution of apoptosis via a common death effector machinery. This article reviews the specific morphological and biochemical hallmarks of apoptosis, the major participating players, and the various signaling pathways involved in triggering this highly complex form of cell death.

Apoptosis↗