Intrastrain variants of herpes simplex virus type 1 isolated from a neonate with fatal disseminated infection differ in the ICP34.5 gene, glycoprotein processing, and neuroinvasiveness.
Humans; Male; Animals; Mice; Amino Acid Sequence; *Genetic Variation; Base Sequence; Molecular Sequence Data; Polymerase Chain Reaction/methods; DNA; Deoxyribonuclease BamHI; Deoxyribonuclease EcoRI; Glycoproteins/metabolism; Viral Envelope Proteins/analysis; Viral Proteins/*genetics; Genes; Viral; Animal; Disease Models; Herpesvirus 1; Inbred BALB C; Amino Acid; Sequence Homology; Sequence Analysis; Nucleic Acid; Polymorphism; *Protein Processing; Post-Translational; Human/*genetics/growth & development/isolation & purification; Restriction Fragment Length
Two intrastrain variants of herpes simplex virus type 1 (HSV-1) were isolated from a newborn with fatal disseminated infection. A small-plaque-producing variant (SP7) was the predominant virus (\textgreater99%) in the brain, and a large-plaque-producing variant (LP5) was the predominant virus (\textgreater99%) in the lung and gastrointestinal tract. EcoRI and BamHI restriction fragment patterns indicated that SP7 and LP5 are related strains. The large-plaque variants produced plaques similar in size to those produced by HSV-1 KOS. Unlike LP5 or KOS, SP7 was highly cell associated and processing of glycoprotein C and glycoprotein D was limited to precursor forms in infected Vero cells. The large-plaque phenotype from KOS could be transferred into SP7 by cotransfection of plasmids containing the EK or JK EcoRI fragment or a 3-kb plasmid with the UL34.5 gene of HSV-1 KOS together with SP7 DNA. PCR analysis using primers from within the ICP34.5 gene indicated differences for SP7, LP5, and KOS. Sequencing data indicated two sets of deletions in the UL34.5 gene that distinguish SP7 from LP5. Both SP7 and LP5 variants were neurovirulent (lethal following intracranial inoculation of young BALB/c mice); however, the LP5 variant was much less able to cause lethal neuroinvasive disease (footpad inoculation) whereas KOS caused no disease. Passage of SP7 selected for viruses (SLP-5 and SLP-10) which were attenuated for lethal neuroinvasive disease, were not cell-associated, and differed in the UL34.5 gene. UL34.5 from SLP-5 or SLP-10 resembled that of KOS. These findings support a role for UL34.5 in promoting virus egress and for neuroinvasive disease.
Bower J R; Mao H; Durishin C; Rozenbom E; Detwiler M; Rempinski D; Karban T L; Rosenthal K S
Journal of virology
1999
1999-05
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
Strain-dependent structural variants of herpes simplex virus type 1 ICP34.5 determine viral plaque size, efficiency of glycoprotein processing, and viral release and neuroinvasive disease potential.
*Genetic Variation; Animals; Cercopithecus aethiops; Cultured; Encephalitis; Genetic; Herpes Simplex/*physiopathology/virology; Herpesvirus 1; Human/*classification/genetics/*pathogenicity; Inbred BALB C; Male; Mice; Recombination; Transfection; Tumor Cells; Vero Cells; Viral Envelope Proteins/metabolism; Viral Plaque Assay; Viral Proteins/*chemistry/*genetics/metabolism
The ability of certain strains of herpes simplex virus type 1 (HSV-1) to cause encephalitis or neuroinvasive disease in the mouse upon peripheral infection is dependent on a combination of activities of specific forms of viral proteins. The importance of specific variants of ICP34.5 to neuroinvasive disease potential and its correlation with small-plaque production, inefficient glycoprotein processing, and virus release were suggested by comparison of ICP34.5 from the SP7 virus, originally obtained from the brain of a neonate with disseminated disease, and the tissue culture-passaged progeny of SP7 (SLP5 and SLP10) and the KOS321 virus. SLP5, SLP10, and KOS321 are attenuated and exhibit a large-plaque phenotype, including efficient glycoprotein processing and viral release. We show that expression of the KOS321 ICP34.5 protein in cells infected with SP7 or ICP34.5 deletion mutants promotes large plaque formation and efficient viral glycoprotein processing, while expression of the SP7 ICP34.5 protein decreases efficiency of viral glycoprotein processing. In addition, a recombinant virus, 4hS1, with the SP7 ICP34.5 gene replacing the KOS321-like ICP34.5 gene in the SLP10a background, rescues the small-plaque phenotype and neuroinvasive disease. The major difference in the ICP34.5 gene product is the number of Pro-Ala-Thr repeats in the middle region of the protein, with 18 for SP7 and 3 for KOS321. Strain-dependent differences in the ICP34.5 protein can therefore alter the tissue culture behavior and the virulence of HSV-1.
Mao Hanwen; Rosenthal Ken S
Journal of virology
2003
2003-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1128/jvi.77.6.3409-3417.2003" target="_blank" rel="noreferrer noopener">10.1128/jvi.77.6.3409-3417.2003</a>
Turnover of histones and histone variants in postnatal rat brain: effects of alcohol exposure.
*2H2O-labeling; *Brain; *Genetic Variation; *Histone; *Mass spectrometry; *Post-translational modifications; *Postnatal alcohol exposure; *Turnover; Acetylation; Animal; Animals; Cell Proliferation; Disease Models; DNA Damage; Epigenesis; Female; Fetal Alcohol Spectrum Disorders/genetics/*metabolism; Genetic; Histones/*genetics/*metabolism; Humans; Post-Translational; Pregnancy; Protein Processing; Proteomics/*methods; Rats; Sprague-Dawley
BACKGROUND: Alcohol consumption during pregnancy is a significant public health problem and can result in a continuum of adverse outcomes to the fetus known as fetal alcohol spectrum disorders (FASD). Subjects with FASD show significant neurological deficits, ranging from microencephaly, neurobehavioral, and mental health problems to poor social adjustment and stress tolerance. Neurons are particularly sensitive to alcohol exposure. The neurotoxic action of alcohol, i.e., through ROS production, induces DNA damage and neuronal cell death by apoptosis. In addition, epigenetics, including DNA methylation, histone posttranslational modifications (PTMs), and non-coding RNA, play an important role in the neuropathology of FASD. However, little is known about the temporal dynamics and kinetics of histones and their PTMs in FASD. RESULTS: We examined the effects of postnatal alcohol exposure (PAE), an animal model of human third-trimester equivalent, on the kinetics of various histone proteins in two distinct brain regions, the frontal cortex, and the hypothalamus, using in vivo (2)H2O-labeling combined with mass spectrometry-based proteomics. We show that histones have long half-lives that are in the order of days. We also show that H3.3 and H2Az histone variants have faster turnovers than canonical histones and that acetylated histones, in general, have a faster turnover than unmodified and methylated histones. Our work is the first to show that PAE induces a differential reduction in turnover rates of histones in both brain regions studied. These alterations in histone turnover were associated with increased DNA damage and decreased cell proliferation in postnatal rat brain. CONCLUSION: Alterations in histone turnover might interfere with histone deposition and chromatin stability, resulting in deregulated cell-specific gene expression and therefore contribute to the development of the neurological disorders associated with FASD. Using in vivo (2)H2O-labeling and mass spectrometry-based proteomics might help in the understanding of histone turnover following alcohol exposure and could be of great importance in enabling researchers to identify novel targets and/or biomarkers for the prevention and management of fetal alcohol spectrum disorders.
Rachdaoui Nadia; Li Ling; Willard Belinda; Kasumov Takhar; Previs Stephen; Sarkar Dipak
Clinical epigenetics
2017
1905-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1186/s13148-017-0416-5" target="_blank" rel="noreferrer noopener">10.1186/s13148-017-0416-5</a>