The herpes simplex virus UL37 protein is phosphorylated in infected cells

162

Information on the extent of virus DNA transcription and translation in infected tissue is crucial to an understanding of herpesvirus latency. To detect low-abundance latent varicella-zoster virus (VZV) transcripts, poly(A) ؉ RNA extracted from latently infected human trigeminal ganglia was enriched for VZV transcripts by hybridization to biotinylated VZV DNA. After hybridization, the RNA-DNA hybrid was isolated by binding to avidin-coated beads and extensively washed, and the RNA was released by heat denaturation. A lambda-based cDNA library was then constructed from the enriched RNA. PCR and DNA sequencing of DNA extracted from the cDNA library revealed the presence of VZV genes 21, 29, 62, and 63, but not VZV genes 4, 10, 40, 51, and 61, in the enriched cDNA library. These findings confirm the detection of VZV gene 29 and 62 transcripts on Northern (RNA) blots prepared from latently infected human ganglia (J.

Section: Discussionmentioning

confidence: 99%

Varicella-zoster virus (VZV) transcription during latency in human ganglia: detection of transcripts mapping to genes 21, 29, 62, and 63 in a cDNA library enriched for VZV RNA

Cohrs

Barbour

Gilden

1996

162

“…5h). HSV UL37 has also been shown to be a phosphoprotein that forms a complex with the DNA-bound HSV ICP8 (1,18). Further analysis of VZV gene 21 protein is needed to determine its kinetic class, whether it is phosphorylated, and if it is associated with VZV ORF 29, a major DNA-binding protein homologous to HSV ICP8.…”

Section: Discussionmentioning

confidence: 99%

Localization of Varicella-Zoster Virus Gene 21 Protein in Virus-Infected Cells in Culture

Mahalingam¹,

Lasher²,

Wellish³

et al. 1998

Although four varicella-zoster virus (VZV) genes have been shown to be transcribed in latently infected human ganglia, their role in the development and maintenance of latency is unknown. To study these VZV transcripts, we decided first to localize their expression products in productively infected cells. We began with VZV gene 21, whose open reading frame (ORF) is 3,113 bp. We cloned the 5′ and 3′ ends and the predicted antigenic segments of the ORF as 1292-, 1280-, and 880-bp DNA fragments, respectively, into the prokaryotic expression vector pGEX-2T. The three VZV 21 ORFs were expressed as approximately 75-, 73-, and 59-kDa glutathione S-transferase fusion proteins in Escherichia coli. To prepare polyclonal antibodies that would recognize all potential epitopes on the VZV gene 21 protein, rabbits were inoculated with a mixture of the three fusion proteins, and antisera were obtained and affinity purified. Immunohistochemical and immunoelectron microscopic analyses using these antibodies revealed VZV ORF 21 protein in both the nucleus and cytoplasm of VZV-infected cells. When these antibodies were applied to purified VZV nucleocapsids, intense staining was seen in their central cores.

“…In the only study to identify latent VZV transcripts by Northern (RNA) blotting, hundreds of human ganglia were pooled and VZV gene 29 and The product of VZV gene 21 has not been identified. However, the HSV type 1 (HSV-1) homolog, UL37, encodes a late (␥1) 120-kDa phosphoprotein which associates with HSV-1 ICP8 (the major DNA-binding protein) in infected cells (1,14,22,23). VZV gene 29 encodes the major DNA-binding protein in VZV-infected cells which, by analogy to HSV-1 ICP8, is an early protein (6,16).…”

Section: Fig 2 Sensitivity Of Detection Of Vzv Gene 21 and 40 Transmentioning

confidence: 99%

Varicella-zoster virus (VZV) transcription during latency in human ganglia: prevalence of VZV gene 21 transcripts in latently infected human ganglia

Cohrs

Barbour

Mahalingam

et al. 1995