Herpes simplex virus 1 tegument protein US11 downmodulates the RLR signaling pathway via direct interaction

On the basis of these observations, we predict that the abilities of UL46 and UL47 to enhance alpha TIF-mediated transcription could result from a stoichiometric association of VP11/12 and VP13/14 with alpha TIF within the infecting virion. With respect to their antiviral effects, the autophagy proteins target viral components or virions for lysosomal degradation in a process termed xenophagy (8), and they also play a role in initiating innate and adaptive immune system responses to viral infections (5, 9). Extensive membrane fusion can be induced by coexpressing glycoproteins gB, gD, and gH/gL in cell lines (25, 26), suggesting that these glycoproteins are sufficient for membrane fusion. Virol. Densitometric analysis of purified virions showed that the levels of VP11/12 and VP13/14 in the virion tegument were near the molar ratios of alpha TIF. 76:3282-3291, 2002), suggests that these associations may be important for the intracellular movement of viral components. This RNA-binding domain (RBD) shows some homology to the Epstein–Barr virus SM protein (12), but it is not homologous to any other known proteins and thus the mechanism by which it recognizes and binds RNA may be novel.

This leads to phosphorylation of the α-subunit of the eukaryotic translation initiation factor (eIF2α) with complete inhibition of translation. This recruitment requires the interaction of HIPK2 with the PML-3 protein (18). The UL11 protein contains 96 amino acids and accumulates on the cytoplasmic faces of both nuclear and Golgi apparatus-derived membranes within infected cells (1, 24). ↵† Present address: Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322. A virus in which the UL11 gene was restored produced wild-type levels of total and extracellular virus and was indistinguishable from wild-type virus upon analysis by electron microscopy. In this membrane fusion model, binding of gD to its cognate receptors, including nectin-1, herpesvirus entry mediator (HVEM), and other receptors (16–22), is thought to trigger sequential conformational changes in gH/gL and gB, causing fusion of the viral envelope with cellular membranes during virus entry as well as fusion among cellular membranes (23, 24). Rather, the observed range in clinical outcomes appears to reflect differences in intrinsic resistance to infection.

Long-term persistence of gene-modified cells could potentially be achieved by administering immunosuppressive regimens commonly used in the prevention of solid organ graft rejection, graft-versus-host disease, and autoimmune disorders. Fax: (421) 624-1401. Conclusions In this population of women at risk for HIV-1, seroprevalence of HSV-2 was high, with potentially important differences by age and site of enrollment. Thus, PACT is a true protein activator of PKR. For example, if the current year is 2008 and a journal has a 5 year moving wall, articles from the year 2002 are available. To test whether the highly structured 5′ untranslated region (5’UTR) of the true-late gC mRNA is the primary obstacle for translation initiation, we replaced it with the less structured 5’UTR of the γ-actin mRNA. Rather, the researchers assert that the diseases must have been transferred, given the timeline and geography of human migration and what pathogen genomes tell us about the ancestry of disease.

To test whether the highly structured 5′ untranslated region (5′UTR) of the true-late gC mRNA is the primary obstacle for translation initiation, we replaced it with the less structured 5′UTR of the γ-actin mRNA. Ablation of this phosphorylation abolished US3-mediated downregulation of CD1d expression, suggesting that phosphorylation of KIF3A is the primary mechanism of HSV-1 suppression of CD1d expression by US3 protein. These observations performed with intron-containing constructs provide evidence that HSV-1 Us11 protein is not directly involved in the cytoplasmic accumulation of viral mRNAs but may be rather acting as an auxiliary protein, thus allowing this retroviral protein to fulfill the nuclear export of these transcripts and to rescue HIV-1 production. The distribution of the phosphate bound to viral polypeptides varied depending on the Mg2+ concentration and pH. Further evidence for the presence of a C-terminal regulatory domain was provided by single-amino-acid substitutions at conserved cysteines (C269S, C271S, and C357S), which enabled the efficient interaction of full-length UL16 with UL11. Intravenous acyclovir (IV ACV) was started in 10/11 cases (range: 3-10 days), switched to valaciclovir (VACV) (range: 5-7 days); one patient was treated with ACV per os for 10 days. In contrast, when sunscreen was applied before UV exposure, no lesions developed, but 1 of the 35 patients shed virus at the exposure site.

The results of these experiments indicate that immunization with gD produces protection against latent ganglionic infection in 56% of the immunized animals, and provides protection against keratitis and death following HSV corneal challenge. Entry depended on viral gD and was diminished in the absence of cellular GAGs. We investigated the CD8 T cell response to HSV-2 in chronically infected individuals by sequencing the hypervariable regions encoding TCR alpha and beta polypeptides from T cell clones recognizing virion protein 22 aa 49-57, an immunodominant epitope.

Herpes Simplex Virus 1 Tegument Protein US11 Downmodulates the RLR Signaling Pathway via Direct Interaction

This process is called secondary envelopment (reviewed in Mettenleiter, 2002). K-bZIP (red) and CDK2 (green) were detected with Alexa Fluor 555-conjugated goat F(ab′)2 anti-rabbit immunoglobulin G and Alexa Fluor 488-conjugated goat F(ab′)2 anti-mouse immunoglobulin G. Data represent means from three independent experiments. A, lane 6). 2003; Pati et al. CT26 tumors were established in mice previously exposed to HSV (see Materials and Methods for details), and the resulting tumors were injected intratumorally with DISC/mGM-CSF. Because claspin was absent, ATR-Chk1 signaling in response to replication stress was interrupted and the intra S phase cell cycle checkpoint was relaxed.

Animals pretreated with the proenkephalin vector were resistant to hyperalgesic sensitization of myelinated and unmyelinated nociceptors. Since fibrillarin is expressed in both the nucleolus and coiled bodies, we chose to use in the present study rabbit antisera against p80-coilin, a structural protein specific for the coiled body, to confirm that these nuclear foci are coiled bodies in double-labeling assays. Fig 4. Moreover, there is no direct homology between the HSV-2 genome and coding sequences in the producer cell. These experiments identified the presence of reactivable virus (Figure 3B) and infectious virions (Figure 3C) only in the IVIS+ group. B. Monolayers were infected at MOIs of 0.1, 1, and 5 PFU/cell in a volume of 100 μl/plate.

The DNA was then purified from the soluble and insoluble chromatin fractions and quantitated. (A) Two KSHV positive cells (BC3 and JSC-1), and a KSHV negative cells (BJAB) were treated with or without different concentration of nocodazole (200 ng/mL, 300 ng/mL, 500 ng/mL or 1 µg/mL) for 24 hours. Recombinant pGEX vectors were grown in E. At the indicated times after infection, the medium was removed, the monolayers were washed twice with cold PBS and scraped into 150 μl of RNA extraction buffer (DirectProtect; Ambion), and the resulting cell extracts were transferred to Eppendorf tubes. 1, lanes 3, 4, 9, and 10). 1A). P3HR1 mRNA does not cross-hybridize to any of the KSHV probes.

(B–D) EGFP and mCherry expression in infected HDFs. Total DNA was subjected to restriction enzyme digestion overnight and electrophoresed on 0.8% agarose gels. The Signal Transduction PathwayFinder cDNA array from Superarray incorporates 96 targets covering 19 associated signalling pathways, in addition to two negative controls corresponding to pUC18 plasmid DNA and a ‘blank’ spot, as well as four positive controls representing the housekeeping genes for GAPDH, beta actin, cyclophilin A and ribosomal protein L13a. The anti-myc monoclonal antibody (MAb) (immunoglobulin G1 [IgG1]) (Invitrogen) was used at a 1:500 dilution. Every gel slice was subjected to in-gel digestion with trypsin overnight at 37°C. Control B6 and B-cell−/− mice did not receive any serum. To start a productive, lytic replication cycle [110, 111], a temporal and sequential cascade of immediate early (IE), early (E) and late (L) gene expression is initiated [104].

Harvested cells were then fixed and permeabilized in 70% ethanol in PBS and stored at −20°C. The dry pellet of nucleic acid was resuspended in 10 mM Tris-HCl-1 mM EDTA. Samples were precleared overnight with protein A-agarose beads (Roche) to minimize background and then reacted with the polyclonal R69 anti-gB antibody (1/800) for 3 h. MS-275 and apicidin, representing two additional classes of HDAC inhibitors, and suberoylanilide hydroxamic acid (SAHA) reactivated EBV in HH514-16 cells; this activity was also inhibited by VPA (29). Viral titers were determined as viral DNA equivalents by quantitative PCR and were confirmed by virus infection in inoculated cells. Viral infections.Viral stocks were diluted in cMEM and adsorbed to monolayers of cells plated the previous day. Reduction in vGPCR by RNA interference also resulted in the reduction in KSHV lytic switch ORF50 gene and protein expression.

LightCycler PCR.A 5-μl aliquot of each extracted specimen was added to 15 μl of a master mixture for amplification. Immediate-early promoters from alpha- and betaherpesviruses can be activated by a virus-encoded virion factor (24). Mounting evidence suggests that phosphorylation is a common regulatory mechanism among replication initiators (reviewed in reference50). Human herpesvirus 6B (HHV-6B), the causative agent of exanthema subitum, infects virtually 100% of individuals in the Western world (8, 10). These results provide the first evidence that the virus plays an active role in down-regulating productive infection during acute infection of sensory neurons. From 42 cellular factors that interacted with pUL34 in yeast, twelve were further evaluated in mammalian cells by co-localization studies using immunofluorescence. Infection with DISC-HSV inhibited tumor cell growth both in vitro and in vivo.

KSHV encodes proteins that deregulate key checkpoints in the signalling pathways governing cell proliferation, which may ultimately contribute to the virus’ oncogenic potential. ↵‡ To whom correspondence should be addressed: Medical Research Council Virology Unit, Church St., Glasgow G11 5JR, Scotland, UK. The interferon (IFN)-mediated antiviral response is a major defense of the host immune system. Title: An investigation of the effect of herpes simplex virus 1 infection on host cell metabolism Authors: Grady, Sarah L.