scholarly journals Herpes Simplex Virus Type 1 Cytoplasmic Envelopment Requires Functional Vps4

2007 ◽  
Vol 81 (14) ◽  
pp. 7380-7387 ◽  
Author(s):  
Colin M. Crump ◽  
Catherine Yates ◽  
Tony Minson

ABSTRACT The assembly and egress of herpesviruses are complex processes that require the budding of viral nucleocapsids into the lumen of cytoplasmic compartments to form mature infectious virus. This envelopment stage shares many characteristics with the formation of luminal vesicles in multivesicular endosomes. Through expression of dominant-negative Vps4, an enzyme that is essential for the formation of luminal vesicles in multivesicular endosomes, we now show that Vps4 function is required for the cytoplasmic envelopment of herpes simplex virus type 1. This is the first example of a large enveloped DNA virus engaging the multivesicular endosome sorting machinery to enable infectious virus production.

2006 ◽  
Vol 87 (12) ◽  
pp. 3483-3494 ◽  
Author(s):  
Sven Hoppe ◽  
Mario Schelhaas ◽  
Verena Jaeger ◽  
Timo Liebig ◽  
Philipp Petermann ◽  
...  

The aim of this study was to understand how molecular determinants of epithelial cells influence initial infection by herpes simplex virus type 1 (HSV-1). Upon infection of the epithelial MDCKII cell line, enhanced association of virus particles with cells forming actin protrusions was observed, suggesting a putative role of actin dynamics in HSV-1 infection. Thus, the impact of the small Rho-like GTPases Rac1, Cdc42 and RhoA acting as key regulators of actin dynamics was addressed. Endogenous Rac1 and Cdc42 were temporarily activated at 15 and 30 min after HSV-1 infection. When constitutively active Cdc42 or Rac1 mutants were expressed transiently, a significant decrease in infectivity was observed, whereas expression of RhoA mutants had no influence. Furthermore, dominant-negative Cdc42 led to decreased infectivity, whereas dominant-negative Rac1 had no effect. So far, the study of potential effectors indicated that Rac1/Cdc42 mutants inhibited infectivity independently of p21-activated kinase (Pak1). The inhibitory effect of Rac1/Cdc42 mutant expression on HSV-1 infection was characterized further and it was found that binding, internalization and transport of HSV-1 were not affected by expression of Rac1/Cdc42 mutants. Thus, these results provide the first evidence for a role of Rac1/Cdc42 signalling during early HSV-1 infection and suggest a mechanism relying on virus-induced regulation of Rac1/Cdc42 activities.


1997 ◽  
Vol 60 (10) ◽  
pp. 1259-1261 ◽  
Author(s):  
D. BARDELL

Droplets of saliva containing herpes simplex virus type 1 were placed on the skin of tomatoes and the upper surface of lettuce leaves. There was no loss of virus infectivity titer at refrigerator temperature (2°C) at any time examined up to 1 h, the longest period tested. At room temperature (22 to 24°C) there was a 2-log drop in titer between 30 and 60 min, but some infectious virus was still present at 1 h. The virus-containing saliva remained in a liquid state at 2°C. At 22 to 24°C the droplets became dry at approximately 50 min. Implications of the findings are discussed.


2009 ◽  
Vol 83 (21) ◽  
pp. 11254-11264 ◽  
Author(s):  
Tobias Pawliczek ◽  
Colin M. Crump

ABSTRACT Herpes simplex virus type 1 (HSV-1) acquires its mature virus envelope by budding into the lumen of cytoplasmic membranous compartments carrying the viral glycoproteins. In a cellular context, a budding process with identical topology occurs during the formation of intraluminal vesicles in multivesicular bodies. The cellular machinery that mediates this budding process is composed of four protein complexes termed endosomal sorting complexes required for transport (ESCRTs) and several associated proteins, including the ATPase VPS4. We have recently shown that functional VPS4 is specifically required for the cytoplasmic envelopment of HSV-1. We now demonstrate that, consistent with a role of VPS4 in virus envelopment, dominant-negative ESCRT-III proteins potently block HSV-1 production. Retroviruses are known to recruit the ESCRT machinery by small peptide motifs termed late domains. These late domains interact with various ESCRT components and thereby promote ESCRT recruitment. The best-characterized late-domain interacting ESCRT proteins are ALIX and TSG101. The presence of potential ALIX and TSG101 binding sequence motifs in various structural HSV-1 proteins suggested a functional role of these proteins in HSV-1 envelopment. We therefore used a set of dominant-negative proteins, as well as RNA interference, to characterize the contribution of ALIX and TSG101 to HSV-1 production. Interestingly, despite the strict requirement for a functional ESCRT-III complex, our data suggest that HSV-1 production is independent of ALIX and TSG101 expression. In line with these data, we also find that ESCRT-III proteins and VPS4A/B are specifically incorporated into mature HSV-1 virions.


1997 ◽  
Vol 8 (4) ◽  
pp. 317-326 ◽  
Author(s):  
AM Thackray ◽  
HJ Field

Mice with or without immunosuppression by cyclosporin (Cy) were inoculated with herpes simplex virus type 1 in the ear pinna. Without immunosuppression, 20% of the mice died; clinical signs resolved in survivors and infectious virus was cleared by 7 to 10 days post-inoculation (p.i.). With Cy, mortality was 50%, clinical signs increased and infectious virus persisted. Mice were treated with either valaciclovir (VACV) or famciclovir (FCV) from days 1–5 or 5–10 p.i. and both compounds moderated the disease, but only FCV led to rapid restoration of body weight and complete protection from mortality. Resolution of clinical signs was more marked with immunosuppression. On cessation of VACV therapy, infectious virus recurred on individual days. Without immunosuppression, recurrence was detected in neural tissues only, but with Cy, infectious virus also recurred in skin. No recurrences of infectious virus were observed in any FCV-treated mice.


2004 ◽  
Vol 78 (11) ◽  
pp. 5756-5765 ◽  
Author(s):  
Hanka Augustinova ◽  
Daniela Hoeller ◽  
Feng Yao

ABSTRACT By selectively regulating the expression of the trans-dominant-negative mutant polypeptide UL9-C535C, of herpes simplex virus type 1 (HSV-1) origin binding protein UL9 with the tetracycline repressor (tetR)-mediated gene switch, we recently generated a novel replication-defective and anti-HSV-specific HSV-1 recombinant, CJ83193. The UL9-C535C peptides expressed by CJ83193 can function as a potent intracellular therapy against its own replication, as well as the replication of wild-type HSV-1 and HSV-2 in coinfected cells. In this report, we demonstrate that CJ83193 cannot initiate acute productive infection in corneas of infected mice nor can it reactivate from trigeminal ganglia of mice latently infected by CJ83193 in a mouse ocular model. Given that CJ83193 is capable of expressing the viral α, β, and γ1 genes but little or no γ2 genes, we tested the vaccine potential of CJ83193 against HSV-1 infection in a mouse ocular model. Our studies showed that immunization with CJ83193 significantly reduced the yields of challenge HSV in the eyes and trigeminal ganglia on days 3, 5, and 7 postchallenge. Like in mice immunized with the wild-type HSV-1 strain KOS, immunization of mice with CJ83193 prevents the development of keratitis and encephalitis induced by corneal challenge with wild-type HSV-1 strain mP. Delayed-type hypersensitivity (DTH) assays demonstrate that CJ83193 can elicit durable cell-mediated immunity at the same level as that of wild-type HSV-1 and is more effective than that induced by d27, an HSV-1 ICP27 deletion mutant. Moreover, mice immunized with CJ83193 developed strong, durable HSV-1-neutralizing antibodies at levels at least twofold higher than those induced by d27. The results presented in this report have shed new light on the development of effective HSV viral vaccines that encode a unique safety mechanism capable of inhibiting the mutant's own replication and that of wild-type virus.


1995 ◽  
Vol 58 (10) ◽  
pp. 1150-1152 ◽  
Author(s):  
D. BARDELL

Droplets of saliva containing herpes simplex virus type 1 were placed on latex disposable gloves. The temperature at the surface of the gloved hand was 34°C. There was no loss of infectious virus before 15 min. Between 15 and 30 min there was a 2-log-cycle drop in titer, and infectious virus could still be recovered after 1 h, the longest period tested. The drop in titer was due to drying of the saliva, which occurred at approximately 21 min. Infectious virus was transferred by touch to lettuce and ham at 0 min when the virus-containing droplets were in a liquid condition, and after 30 and 60 min when the droplets were dry.


2002 ◽  
Vol 76 (17) ◽  
pp. 8939-8952 ◽  
Author(s):  
Ashley E. Reynolds ◽  
Elizabeth G. Wills ◽  
Richard J. Roller ◽  
Brent J. Ryckman ◽  
Joel D. Baines

ABSTRACT The wild-type UL31, UL34, and US3 proteins localized on nuclear membranes and perinuclear virions; the US3 protein was also on cytoplasmic membranes and extranuclear virions. The UL31 and UL34 proteins were not detected in extracellular virions. US3 deletion caused (i) virion accumulation in nuclear membrane invaginations, (ii) delayed virus production onset, and (iii) reduced peak virus titers. These data support the herpes simplex virus type 1 deenvelopment-reenvelopment model of virion egress and suggest that the US3 protein plays an important, but nonessential, role in the egress pathway.


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