Characterization of Extracellular HSV-1 Virions by Proteomics

2014 ◽  
pp. 181-190
Author(s):  
Roger Lippé
Keyword(s):  
Hsv 1

scholarly journals Characterization of a Novel Alphaherpesvirus Isolated from the Fruit Bat Pteropus lylei in Vietnam

2020 ◽  
Vol 94 (18) ◽  
Author(s):  
Takuya Inagaki ◽  
Souichi Yamada ◽  
Hikaru Fujii ◽  
Tomoki Yoshikawa ◽  
Miho Shibamura ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Lung Fibroblasts ◽  
Whole Genome Sequence ◽  
Replication Capacity ◽  
Human Embryonic Lung ◽  
Host Animal ◽  
Entire Genome ◽  
Fruit Bat ◽  
Hsv 1

ABSTRACT Herpesviruses exist in nature within each host animal. Ten herpesviruses have been isolated from bats and their biological properties reported. A novel bat alphaherpesvirus, which we propose to name “Pteropus lylei-associated alphaherpesvirus (PLAHV),” was isolated from urine of the fruit bat Pteropus lylei in Vietnam and characterized. The entire genome sequence was determined to be 144,008 bp in length and predicted to include 72 genes. PLAHV was assigned to genus Simplexvirus with other bat alphaherpesviruses isolated from pteropodid bats in Southeast Asia and Africa. The replication capacity of PLAHV in several cells was evaluated in comparison with that of herpes simplex virus 1 (HSV-1). PLAHV replicated better in the bat-originated cell line and less in human embryonic lung fibroblasts than HSV-1 did. PLAHV was serologically related to another bat alphaherpesvirus, Pteropodid alphaherpesvirus 1 (PtAHV1), isolated from a Pteropus hypomelanus-related bat captured in Indonesia, but not with HSV-1. PLAHV caused lethal infection in mice. PLAHV was as susceptible to acyclovir as HSV-1 was. Characterization of this new member of bat alphaherpesviruses, PLAHV, expands the knowledge on bat-associated alphaherpesvirology. IMPORTANCE A novel bat alphaherpesvirus, Pteropus lylei-associated alphaherpesvirus (PLAHV), was isolated from urine of the fruit bat Pteropus lylei in Vietnam. The whole-genome sequence was determined and was predicted to include 72 open reading frames in the 144,008-bp genome. PLAHV is circulating in a species of fruit bats, Pteropus lylei, in Asia. This study expands the knowledge on bat-associated alphaherpesvirology.

scholarly journals Longitudinal transcriptomic characterization of viral genes in HSV-1 infected tree shrew trigeminal ganglia

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Erlin Wang ◽  
Yunshuang Ye ◽  
Ke Zhang ◽  
Jinlong Yang ◽  
Daohua Gong ◽  
...  
Keyword(s):  
High Efficiency ◽  
Tree Shrew ◽  
Acute Infection ◽  
Human Infection ◽  
Trigeminal Ganglia ◽  
Tree Shrews ◽  
Infected Tree ◽  
Viral Genes ◽  
Hsv 1

Abstract Background Following acute infection, Herpes Simplex virus-1 (HSV-1) establishes lifelong latency and recurrent reactivation in the sensory neurons of trigeminal ganglia (TG). Infected tree shrew differs from mouse and show characteristics similar to human infection. A detailed transcriptomic analysis of the tree shrew model could provide mechanistic insights into HSV-1 infection in humans. Methods We sequenced the transcriptome of infected TGs from tree shrews and mice, and 4 human donors, then examined viral genes expression up to 58 days in infected TGs from mouse and tree shrew, and compare the latency data with that in human TGs. Results Here, we found that all HSV-1 genes could be detected in mouse TGs during acute infection, but 22 viral genes necessary for viral transcription, replication and viral maturation were not expressed in tree shrew TGs during this stage. Importantly, during latency, we found that LAT could be detected both in mouse and tree shrew, but the latter also has an ICP0 transcript signal absent in mouse but present in human samples. Importantly, we observed that infected human and tree shrew TGs have a more similar LAT region transcription peak. More importantly, we observed that HSV-1 spontaneously reactivates from latently infected tree shrews with relatively high efficiency. Conclusions These results represent the first longitudinal transcriptomic characterization of HSV-1 infection in during acute, latency and recurrent phases, and revealed that tree shrew infection has important similar features with human infection.

scholarly journals Dataset supporting the proteomic characterization of human corneal epithelial cells with HSV-1 infection

Data in Brief ◽  
2019 ◽  
Vol 27 ◽  
pp. 104579
Author(s):  
Zhi-Yi Xu ◽  
Jia-Hui Li ◽  
Mei-Jun Li ◽  
Wen-Lin Zheng ◽  
Hong-Wei Pan
Keyword(s):  
Epithelial Cells ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Human Corneal Epithelial Cells ◽  
Hsv 1

scholarly journals Characterization of Herpes Simplex Viruses Selected in Culture for Resistance to Penciclovir or Acyclovir

2001 ◽  
Vol 75 (4) ◽  
pp. 1761-1769 ◽  
Author(s):  
Robert T. Sarisky ◽  
Matthew R. Quail ◽  
Philip E. Clark ◽  
Tammy T. Nguyen ◽  
Wendy S. Halsey ◽  
...  
Keyword(s):  
Herpes Simplex ◽  
Sequence Similarity ◽  
Cross Resistance ◽  
Phenotypic Analysis ◽  
Herpes Simplex Viruses ◽  
Infected Cells ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Penciclovir (PCV), an antiherpesvirus agent in the same class as acyclovir (ACV), is phosphorylated in herpes simplex virus (HSV)-infected cells by the viral thymidine kinase (TK). Resistance to ACV has been mapped to mutations within either the TK or the DNA polymerase gene. An identical activation pathway, the similarity in mode of action, and the invariant cross-resistance of TK-negative mutants argue that the mechanisms of resistance to PCV and ACV are likely to be analogous. A total of 48 HSV type 1 (HSV-1) and HSV-2 isolates were selected after passage in the presence of increasing concentrations of PCV or ACV in MRC-5 cells. Phenotypic analysis suggested these isolates were deficient in TK activity. Moreover, sequencing of the TK genes from ACV-selected mutants identified two homopolymeric G-C nucleotide stretches as putative hot spots, thereby confirming previous reports examining Acvr clinical isolates. Surprisingly, mutations identified in PCV-selected mutants were generally not in these regions but distributed throughout the TK gene and at similar frequencies of occurrence within A-T or G-C nucleotides, regardless of virus type. Furthermore, HSV-1 isolates selected in the presence of ACV commonly included frameshift mutations, while PCV-selected HSV-1 mutants contained mostly nonconservative amino acid changes. Data from this panel of laboratory isolates show that Pcvr mutants share cross-resistance and only limited sequence similarity with HSV mutants identified following ACV selection. Subtle differences between PCV and ACV in the interaction with viral TK or polymerase may account for the different spectra of genotypes observed for the two sets of mutants.

scholarly journals Characterization of the DNA polymerase gene of varicella-zoster viruses resistant to acyclovir

2001 ◽  
Vol 82 (11) ◽  
pp. 2761-2765 ◽  
Author(s):  
Tomoko Kamiyama ◽  
Masahiko Kurokawa ◽  
Kimiyasu Shiraki
Keyword(s):  
Amino Acid ◽  
Dna Polymerase ◽  
Sequence Similarity ◽  
Polymerase Gene ◽  
Varicella Zoster ◽  
Dna Polymerase Gene ◽  
Simplex Virus ◽  
Polymerase Mutants ◽  
Hsv 1

The nucleotide changes of the DNA polymerase gene and the susceptibility of acyclovir (ACV)-resistant varicella-zoster virus (VZV) mutants to anti-herpetic drugs were determined and compared to those of herpes simplex virus type 1 (HSV-1) mutants. The seven ACV-resistant VZV mutants were classified into three groups, N779S, G805C and V855M, according to the sequences of their DNA polymerase genes. The amino acid substitutions N779S and G805C were identical in position to the N815S and G814C mutations in the HSV-1 DNA polymerase mutants, respectively, and the V855M amino acid substitution was similar to the HSV-1 V892M mutation. All three groups of VZV mutants were susceptible to ACV, phosphonoacetic acid, vidarabine and aphidicolin, at levels similar to those seen with the respective HSV-1 mutants, except for subtle differences that were due possibly to the non-conserved regions in their sequences. Although both the HSV-1 and the VZV DNA polymerase genes show 53% sequence similarity, both viruses essentially show a similar biochemical behaviour.

scholarly journals Characterization of Pseudorabies Virus (PrV) Cleavage-Encapsidation Proteins and Functional Complementation of PrV pUL32 by the Homologous Protein of Herpes Simplex Virus Type 1

2009 ◽  
Vol 83 (8) ◽  
pp. 3930-3943 ◽  
Author(s):  
Walter Fuchs ◽  
Barbara G. Klupp ◽  
Harald Granzow ◽  
Tobias Leege ◽  
Thomas C. Mettenleiter
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Herpes Simplex Virus Type ◽  
Pseudorabies Virus ◽  
Gene Products ◽  
Homologous Protein ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Cleavage and encapsidation of newly replicated herpes simplex virus type 1 (HSV-1) DNA requires several essential viral gene products that are conserved in sequence within the Herpesviridae. However, conservation of function has not been analyzed in greater detail. For functional characterization of the UL6, UL15, UL28, UL32, and UL33 gene products of pseudorabies virus (PrV), the respective deletion mutants were generated by mutagenesis of the virus genome cloned as a bacterial artificial chromosome (BAC) in Escherichia coli and propagated in transgenic rabbit kidney cells lines expressing the deleted genes. Neither of the PrV mutants was able to produce plaques or infectious progeny in noncomplementing cells. DNA analyses revealed that the viral genomes were replicated but not cleaved into monomers. By electron microscopy, only scaffold-containing immature but not DNA-containing mature capsids were detected in the nuclei of noncomplementing cells infected with either of the mutants. Remarkably, primary envelopment of empty capsids at the nuclear membrane occasionally occurred, and enveloped tegument-containing light particles were formed in the cytoplasm and released into the extracellular space. Immunofluorescence analyses with monospecific antisera of cells transfected with the respective expression plasmids indicated that pUL6, pUL15, and pUL32 were able to enter the nucleus. In contrast, pUL28 and pUL33 were predominantly found in the cytoplasm. Only pUL6 could be unequivocally identified and localized in PrV-infected cells and in purified virions, whereas the low abundance or immunogenicity of the other proteins hampered similar studies. Yeast two-hybrid analyses revealed physical interactions between the PrV pUL15, pUL28, and pUL33 proteins, indicating that, as in HSV-1, a tripartite protein complex might catalyze cleavage and encapsidation of viral DNA. Whereas the pUL6 protein is supposed to form the portal for DNA entry into the capsid, the precise role of the UL32 gene product during this process remains to be elucidated. Interestingly, the defect of UL32-negative PrV could be completely corrected in trans by the homologous protein of HSV-1, demonstrating similar functions. However, trans-complementation of UL32-negative HSV-1 by the PrV protein was not observed.

Characterization of the nuclear import mechanisms of HSV-1 UL31

2016 ◽  
Vol 397 (6) ◽  
pp. 555-561 ◽  
Author(s):  
Mingsheng Cai ◽  
Jiang Si ◽  
Xiaowei Li ◽  
Zhancheng Zeng ◽  
Meili Li
Keyword(s):  
Nuclear Import ◽  
Nuclear Translocation ◽  
Fluorescent Microscopy ◽  
Multiple Roles ◽  
Live Cells ◽  
Biological Functions ◽  
Herpes Simplex Virus 1 ◽  
Simplex Virus ◽  
Hsv 1

Abstract As an important protein, UL31 has been demonstrated to play multiple roles in herpes simplex virus 1 (HSV-1) replication. Previous studies showed that UL31 predominantly locates in the nucleus in chemical fixed cells and live cells, however, the determining mechanisms for its nuclear translocation is not clear. In the present study, by utilizing live cells fluorescent microscopy and co-immunoprecipitation assays, the nuclear import of UL31 was characterized to be dependent on Ran-, importin α1- and transportin-1-mediated pathway. Therefore, these results will promote the understanding of UL31-mediated biological functions in HSV-1 infection cycle.

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