Recruitment of Herpes Simplex Virus Type 1 Transcriptional Regulatory Protein ICP4 into Foci Juxtaposed to ND10 in Live, Infected Cells

Everett, Roger D.; Sourvinos, George; Orr, Anne

doi:10.1128/jvi.77.6.3680-3689.2003

Cited by 96 publications

(134 citation statements)

References 45 publications

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“…Therefore, we tested whether an ICP0-null mutant virus could be complemented to infect HFFF-2 cells as efficiently as HSV-1 strain 17. Virus dl0C4 (see Materials and Methods) contains the ICP0 deletion mutation of dl1403 and the ECFP-linked ICP4 fusion protein of vECFP-ICP4 (19). The ECFP fusion moiety has no detectable effect on ICP4 functionality (19), and virus dl0C4 behaved in a manner very similar to that of dl1403 in all assays tested, including analysis by Western blotting, growth curves, and titration of PFU and FFU in Vero, HFFF-2, and U2OS cells (data not shown).…”

Section: Resultsmentioning

confidence: 99%

Phenotype of a Herpes Simplex Virus Type 1 Mutant That Fails To Express Immediate-Early Regulatory Protein ICP0

Everett¹,

Boutell²,

Orr³

2004

J Virol

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125

194

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Herpes simplex virus type 1 (HSV-1) immediate-early (IE) regulatory protein ICP0 is required for efficient progression of infected cells into productive lytic infection, especially in low-multiplicity infections of limitedpassage human fibroblasts. We have used single-cell-based assays that allow detailed analysis of the ICP0-null phenotype in low-multiplicity infections of restrictive cell types. The major conclusions are as follows: (i) there is a threshold input multiplicity above which the mutant virus replicates normally; (ii) individual cells infected below the threshold multiplicity have a high probability of establishing a nonproductive infection; (iii) such nonproductively infected cells have a high probability of expressing IE products at 6 h postinfection; (iv) even at 24 h postinfection, IE protein-positive nonproductively infected human fibroblast cells exceed the number of cells that lead to plaque formation by up to 2 orders of magnitude; (v) expression of individual IE proteins in a proportion of the nonproductively infected cells is incompletely coordinated; (vi) the nonproductive cells can also express early gene products at low frequencies and in a stochastic manner; and (vii) significant numbers of human fibroblast cells infected at low multiplicity by an ICP0-deficient virus are lost through cell death. We propose that in the absence of ICP0 expression, HSV-1 infected human fibroblasts can undergo a great variety of fates, including quiescence, stalled infection at a variety of different stages, cell death, and, for a minor population, initiation of formation of a plaque.

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Section: Resultsmentioning

confidence: 99%

Phenotype of a Herpes Simplex Virus Type 1 Mutant That Fails To Express Immediate-Early Regulatory Protein ICP0

Everett¹,

Boutell²,

Orr³

2004

J Virol

Self Cite

125

194

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“…Previously published data suggest that concurrent with their dissolution the ND10 bodies evolve into replication compartments in which viral DNA is synthesized with the participation of a set of viral proteins that include ICP8 and packaged into capsids (9,52,53). In early stages, they appear as small dense bodies (Fig.…”

Section: Viral Replication Is Suppressed In Sp100 −/− Cells Pretreatementioning

confidence: 99%

“…Viral DNA synthesis and some early stages of virion assembly occur in nuclear structures and, at least in the initial stages of formation, at ND10 bodies (9,52,53). In this series of experiments, we used immunostaining with an antibody to ICP8 to visualize the presence of replication compartments.…”

Section: Formation Of Viral Replication Compartments Is Hampered In Thementioning

confidence: 99%

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The SP100 component of ND10 enhances accumulation of PML and suppresses replication and the assembly of HSV replication compartments

Roizman

2017

Proc. Natl. Acad. Sci. U.S.A.

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Nuclear domain 10 (ND10) bodies are small (0.1–1 μM) nuclear structures containing both constant [e.g., promyelocytic leukemia protein (PML), SP100, death domain-associated protein (Daxx)] and variable proteins, depending on the function of the cells or the stress to which they are exposed. In herpes simplex virus (HSV)-infected cells, ND10 bodies assemble at the sites of DNA entering the nucleus after infection. In sequence, the ND10 bodies become viral replication compartments, and ICP0, a viral E3 ligase, degrades both PML and SP100. The amounts of PML and SP100 and the number of ND10 structures increase in cells exposed to IFN-β. Earlier studies have shown that PML has three key functions. Thus, (i) the interaction of PML with viral components facilitates the initiation of replication compartments, (ii) viral replication is significantly less affected by IFN-β in PML−/− cells than in parental PML+/+ cells, and (iii) viral yields are significantly lower in PML−/− cells exposed to low ratios of virus per cell compared with parental PML+/+ cells. This report focuses on the function of SP100. In contrast to PML−/− cells, SP100−/− cells retain the sensitivity of parental SP100+/+ cells to IFN-β and support replication of the ΔICP0 virus. At low multiplicities of infection, wild-type virus yields are higher in SP100−/− cells than in parental HEp-2 cells. In addition, the number of viral replication compartments is significantly higher in SP100−/− cells than in parental SP100+/+ cells or in PML−/− cells.

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“…Western blotting and chemiluminescence detection were performed as previously described. 87 Real-time PCR The Nanogen Q-CMV Real-Time PCR kit (Nanogen Advanced Diagnostics,Italy) was used to quantify the HCMV genomes in combination with the ABI 7500 Fast system (ABI). The primers for HCMV are specific for the exon 4 region of the HCMV MIEA gene (major immediate early antigen, HCMV UL123) while in parallel, a region of the human b globin gene is also amplified.…”

Section: Antibodies/immunofluorescence/western Blot Analysismentioning

confidence: 99%

RhoB is a component of the human cytomegalovirus assembly complex and is required for efficient viral production

et al. 2015

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Human Cytomegalovirus (HCMV), an ubiquitous b-herpesvirus, is a significant pathogen that causes medically severe diseases in immunocompromised individuals and in congenitally infected neonates. RhoB belongs to the family of Rho GTPases, which regulates diverse cellular processes. Rho proteins are implicated in the entry and egress from the host cell of mainly a-and g-herpesviruses, whereas b-herpesviruses are the least studied in this regard. Here, we studied the role of RhoB GTPase during HCMV lytic infection. Microscopy analysis, both in fixed and live infected cells showed that RhoB was translocated to the assembly complex/compartment (AC) of HCMV, a cytoplasmic zone in infected cells where many viral structural proteins are known to accumulate and assembly of new virions takes place. Furthermore, RhoB was localized at the AC even when the expression of the late HCMV AC proteins was inhibited. At the very late stages of infection, cellular projections were formed containing RhoB and HCMV virions, potentially contributing to the successful viral spread. Interestingly, the knockdown of RhoB in HCMV-infected cells resulted in a significant reduction of the virus titer and could also affect the accumulation of AC viral proteins at this subcellular compartment. RhoB knockdown also affected actin fibers' structure. Actin reorganization was observed at late stages of infection originating from the viral AC and surrounding the cellular projections, implying a potential interplay between RhoB and actin during HCMV assembly and egress. In conclusion, our results demonstrate for the first time that RhoB is a constituent of the viral AC and is required for HCMV productive infection.

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Recruitment of Herpes Simplex Virus Type 1 Transcriptional Regulatory Protein ICP4 into Foci Juxtaposed to ND10 in Live, Infected Cells

Cited by 96 publications

References 45 publications

Phenotype of a Herpes Simplex Virus Type 1 Mutant That Fails To Express Immediate-Early Regulatory Protein ICP0

Phenotype of a Herpes Simplex Virus Type 1 Mutant That Fails To Express Immediate-Early Regulatory Protein ICP0

The SP100 component of ND10 enhances accumulation of PML and suppresses replication and the assembly of HSV replication compartments

RhoB is a component of the human cytomegalovirus assembly complex and is required for efficient viral production

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