Small Dense Nuclear Bodies Are the Site of Localization of Herpes Simplex Virus 1 U
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            3 and U
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            4 Proteins and of ICP22 Only When the Latter Protein Is Present

Markovitz, Nancy S.; Roizman, Bernard

doi:10.1128/jvi.74.1.523-528.2000

Cited by 29 publications

(26 citation statements)

References 23 publications

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“…11). The nuclear localization of ICP22 was also characterized by expression in small, dense nuclear structures (14,30), whereas this pattern was observed in only a few VZV-infected cells during replication in melanoma cells.…”

Section: Downloaded Frommentioning

confidence: 99%

The Immediate-Early 63 Protein of Varicella-Zoster Virus: Analysis of Functional Domains Required for Replication In Vitro and for T-Cell and Skin Tropism in the SCIDhu Model In Vivo

Baiker¹,

Bagowski²,

Ito³

et al. 2004

J Virol

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The immediate-early 63-kDa (IE63) protein of varicella-zoster virus (VZV) is a phosphoprotein encoded by open reading frame (ORF) ORF63/ORF70. To identify functional domains, 22 ORF63 mutations were evaluated for effects on IE63 binding to the major VZV transactivator, IE62, and on IE63 phosphorylation and nuclear localization in transient transfections, and after insertion into the viral genome with VZV cosmids. The IE62 binding site was mapped to IE63 amino acids 55 to 67, with R59/L60 being critical residues. Alanine substitutions within the IE63 center region showed that S165, S173, and S185 were phosphorylated by cellular kinases. Four mutations that changed two putative nuclear localization signal (NLS) sequences altered IE63 distribution to a cytoplasmic/nuclear pattern. Only three of 22 mutations in ORF63 were compatible with recovery of infectious VZV from our cosmids, but infectivity was restored by inserting intact ORF63 into each mutated cosmid. The viable IE63 mutants had a single alanine substitution, altering T171, S181, or S185. These mutants, rOKA/ORF63rev[T171], rOKA/ORF63rev[S181], and rOKA/ORF63rev[S185], produced less infectious virus and had a decreased plaque phenotype in vitro. ORF47 kinase protein and glycoprotein E (gE) synthesis was reduced, indicating that IE63 contributed to optimal expression of early and late gene products. The three IE63 mutants replicated in skin xenografts in the SCIDhu mouse model, but virulence was markedly attenuated. In contrast, infectivity in T-cell xenografts was not altered. Comparative analysis suggested that IE63 resembled the herpes simplex virus type 1 U S 1.5 protein, which is expressed colinearly with ICP22 (U S 1). In summary, most mutations of ORF63 made with our VZV cosmid system were lethal for infectivity. The few IE63 changes that were tolerated resulted in VZV mutants with an impaired capacity to replicate in vitro. However, the IE63 mutants were attenuated in skin but not T cells in vivo, indicating that the contribution of the IE63 tegument/regulatory protein to VZV pathogenesis depends upon the differentiated human cell type which is targeted for infection within the intact tissue microenvironment.Varicella-zoster virus (VZV), a member of the alphaherpesvirus subfamily of the Herpesviridae, causes varicella during primary infection and herpes zoster when it reactivates from latency in sensory ganglia. The VZV genome is a doublestranded DNA molecule of approximately 125 kb with open reading frames (ORFs) that are known or predicted to encode at least 70 distinct gene products (1). The genome consists of two main coding regions, the unique long (U L ) and unique short (U S ) regions, each of which is flanked by internal repeat (IR) and terminal repeat (TR) sequences. Three ORFs, including ORF63/70 as well as ORF62/71 and ORF64/69, are located within the repeat sequences and are therefore duplicated within the VZV genome (7).The development of VZV cosmid systems has permitted the functional analysis of VZV gene products by deleting ORFs or i...

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Section: Downloaded Frommentioning

confidence: 99%

The Immediate-Early 63 Protein of Varicella-Zoster Virus: Analysis of Functional Domains Required for Replication In Vitro and for T-Cell and Skin Tropism in the SCIDhu Model In Vivo

Baiker¹,

Bagowski²,

Ito³

et al. 2004

J Virol

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“…During later infection, ICP22 aggregates with UL3, UL4, and UL20.5 proteins as discrete, dense spots in the nucleus (11,18,27). However, deletion of ICP22 resulted in a decreased accumulation of UL3 and UL4, and neither UL3 nor UL4 localizes to…”

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confidence: 99%

“…However, deletion of ICP22 resulted in a decreased accumulation of UL3 and UL4, and neither UL3 nor UL4 localizes to the dense nuclear structures. Furthermore, ICP22 by itself is sufficient to form small, dense nuclear bodies in transfection assays without other viral proteins (18). Consequently, it was suggested that ICP22 may direct UL3 and UL4 to the dense nuclear structure (18).…”

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confidence: 99%

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Comprehensive Characterization of Interaction Complexes of Herpes Simplex Virus Type 1 ICP22, UL3, UL4, and UL20.5

Xing

Wang

Lin

et al. 2011

J Virol

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It has been reported that herpes simplex virus type 1 UL3, UL4, and UL20.5 proteins are localized to small, dense nuclear bodies together with ICP22 in infected cells. In the present study, we comprehensively characterized these interactions by subcellular colocalization, coimmunoprecipitation, and bimolecular fluorescence complementation assays. For the first time, it was demonstrated that both UL3 and UL20.5 are targeted to small, dense nuclear bodies by a direct interaction with ICP22, whereas UL4 colocalizes with ICP22 through its interaction with UL3 but not UL20.5 or ICP22. There was no detectable interaction between UL3 and UL20.5.Of the 84 open reading frames (ORFs) of herpes simplex virus 1 (HSV-1) known to be expressed, more than half are functionally poorly understood and identified as encoding dispensable proteins for viral replication at least in some cell lines (21,22). However, these ORFs appear to be essential for viral survival in nature, since viruses lacking these genes have not been isolated. Because mutant viruses lacking these genes have no obvious phenotype in infected cells, it remains difficult to functionally characterize these genes.As an immediate-early protein, ICP22 appears to be essential for viral replication (5, 19). It localizes in the nucleus of infected cells and functions as a transcriptional repressor for both viral and cellular genes (5). Furthermore, ICP22 associates with transcriptional complexes containing the viral transactivator protein ICP4, RNA polymerase II (4, 9, 14, 16), and other host factors, including cdc25C (25, 26) and cdk9 (7). Previous studies have reported that both UL3 and UL4 are expressed late in infection and are dispensable for viral replication in cultured cells in vitro (2,8,12). As a novel identified ORF situated between genes encoding UL20 and UL21 of the HSV-1 genome, the UL20.5 gene is not essential for growth and belongs to the ␥2 kinetic class (27). Nevertheless, although the proteins encoded by these genes are dispensable for growth in cell culture, their functions are still unknown.ICP22 localizes to small, dense nuclear structures early in infection, followed by the transition to the more diffuse replication complexes with nascent DNA and RNA polymerase II, ICP4, and other proteins associated with late gene transcription after the onset of viral DNA synthesis (11,14). During later infection, ICP22 aggregates with UL3, UL4, and UL20.5 proteins as discrete, dense spots in the nucleus (11,18,27). However, deletion of ICP22 resulted in a decreased accumulation of UL3 and UL4, and neither UL3 nor UL4 localizes to

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“…Products of these latter genes are responsible for the posttranslational phosphorylation of ICP22 protein [41].In 2000, another complex formation was observed, called small dense nuclear bodies, which appear earlier than the RNAPII complex and contains ICP22, UL3, and UL4 [42]. ICP22 and UL13 mediate the phosphorylation of carboxyl terminal domain of RNAPII [43].…”

Section: Genestranscriptionmentioning

confidence: 99%

Effect of us1 gene mutation on pseudorabies virus gene expression

Takács¹

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Small Dense Nuclear Bodies Are the Site of Localization of Herpes Simplex Virus 1 U _L 3 and U _L 4 Proteins and of ICP22 Only When the Latter Protein Is Present

Cited by 29 publications

References 23 publications

The Immediate-Early 63 Protein of Varicella-Zoster Virus: Analysis of Functional Domains Required for Replication In Vitro and for T-Cell and Skin Tropism in the SCIDhu Model In Vivo

The Immediate-Early 63 Protein of Varicella-Zoster Virus: Analysis of Functional Domains Required for Replication In Vitro and for T-Cell and Skin Tropism in the SCIDhu Model In Vivo

Comprehensive Characterization of Interaction Complexes of Herpes Simplex Virus Type 1 ICP22, UL3, UL4, and UL20.5

Effect of us1 gene mutation on pseudorabies virus gene expression

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Small Dense Nuclear Bodies Are the Site of Localization of Herpes Simplex Virus 1 U L 3 and U L 4 Proteins and of ICP22 Only When the Latter Protein Is Present

Cited by 29 publications

References 23 publications

The Immediate-Early 63 Protein of Varicella-Zoster Virus: Analysis of Functional Domains Required for Replication In Vitro and for T-Cell and Skin Tropism in the SCIDhu Model In Vivo

The Immediate-Early 63 Protein of Varicella-Zoster Virus: Analysis of Functional Domains Required for Replication In Vitro and for T-Cell and Skin Tropism in the SCIDhu Model In Vivo

Comprehensive Characterization of Interaction Complexes of Herpes Simplex Virus Type 1 ICP22, UL3, UL4, and UL20.5

Effect of us1 gene mutation on pseudorabies virus gene expression

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Small Dense Nuclear Bodies Are the Site of Localization of Herpes Simplex Virus 1 U _L 3 and U _L 4 Proteins and of ICP22 Only When the Latter Protein Is Present