The HSV-1 protein ICP34.5 interacts with the cellular phosphatase PP1α to mediate the dephosphorylation of eIF2α and thus antagonizes the PKR signaling pathway () (20). Recently, we utilized mutant viruses lacking one or more viral genes to show that the deletion of either the gK or UL20 gene produced significantly greater defects in virion envelopment and overall virus replication than deletion of the carboxyl terminus of either gD, UL11, gM, or gE alone or in various combinations (44). Further analysis of Us11 mutants revealed that Us11 is also required for maximal rates of translation when expressed in its natural context late in infection (M. Pearson from pMAL-c2X [New England Biolabs (NEB)] to contain a PreScission Protease site inserted into the XmnI restriction site, between sequences encoding maltose-binding protein (MBP) and its fusion partner (in this case, US11) (18). The objective of this study was to discover whether US11 had the potential to bind any other transcripts from HSV-infected cells that would be relevant to its role in the virus lytic cycle. Later during infection, the US11 protein accumulates into nucleoli and is also found in RNP fibrils as well as in clusters of interchromatin granules. For the PRV construct, the UL11 gene was amplified from the PRV Becker genome with a forward primer (5′-GCCTGCGAGATCTCCGTGCTGCTGATCGTCACG-3′) complementary to the sequence 100 bp upstream of the start codon and a reverse primer (5′-CAAACGGGTTTATTGGAATTCGTACGCCCGCGAG-3′) complementary to the 3′ end of the UL11 gene.
HSV-1 UL11 was recently shown to form a protein complex with gE, UL16, and UL21 that may be required for efficient virus spread (43). To test this hypothesis, we used CD11c-diphtheria toxin (DT) receptor-green fluorescent protein (GFP)-transgenic (Tg) mice, generated on a C57BL/6 (B6) background, in which DCs can be transiently depleted in vivo by treatment with low doses of DT (15). Cell lines and viruses.The retroviral packaging cell lines PE501, PA317, and PG13 were grown in Dulbecco’s modified Eagle medium with 10% heat-inactivated fetal bovine serum (FBS; HyClone Laboratories, Logan, Utah) (41, 42). This segment of Us11 localizes to the nucleolus, associates with polysomes, and contains a region important for interaction with PKR in an RNA-dependent manner (5, 27, 39, 40). Translation of most transcripts is cap dependent and initiated by the cap-binding complex eIF4F (composed of eIF4E, eIF4G, eIF4A, eIF4B, and eIF4H) binding to the mRNA 5′ m7G-cap via its eIF4E subunit. Once there, CD1d cycles between the cell surface and endosomal compartment through multiple rounds of endocytosis and recycling steps while surveying lipid antigens for NKT cell recognition (3, 9, 15). UL16 is a 373-aa tegument protein that is also conserved among all herpesviruses (24, 34, 48, 55, 58, 73).
Using a murine model of acute hematogenous HSV-1 infection, we previously reported that viral neuroinvasion was reduced in mice lacking ApoE compared to wild-type mice and that the ApoE dose was directly linked to the HSV-1 DNA concentration detected in the nervous system (8). ↵† Supplemental material for this article may be found at http://jvi.asm.org/. Recognized vehicles of transmission are saliva, semen, cervical fluid, or vesicle fluid from active lesions (63). LK Gilbert et al. Phone: (415) 476-4419. The infant showed normal growth and development at 4 years of age. These findings have implications on the role of BER in viral genome maintenance during lytic replication and reactivation from latency.
Weller, J. Thus, both MLH1 and MSH2 are required but appear to participate in distinct events in the virus life cycle. Mailing address: Department of Microbiology, University of Minnesota Medical School, MMC 196, 420 Delaware St. Eight vidarabine-treated patients (22 percent) and three acyclovir-treated patients (9 percent) had moderate debility. In these cells, PKR is activated but eIF-2α is not phosphorylated, and the phosphatase is not redirected to dephosphorylate eIF-2α. The carboxyl-terminal half of the protein consists of a regular tripeptide repeat of the sequence RXP and constitutes a completely novel RNA-binding domain. Laboratory abnormalities at diagnosis associated with mortality were high creatinine, low platelet counts, prolonged partial thromboplastin time, and a high percentage of band forms on the blood smear.
We undertook genetic approaches to address these issues by infecting two different dominant negative TGF-β receptor type II transgenic mouse lines. RESULTS: There were 16 girls and 16 boys, aged 9-44 months (median age, 19 months). Recent studies using knockout (KO) mice demonstrated that the gD receptors herpesvirus entry mediator (HVEM) and nectin-1 are the primary entry receptors for HSV-2 in the mouse vagina and brain. Further, in vitro studies demonstrated that a recombinant expressed US11 protein binds PKR, blocks the phosphorylation of the α subunit of eukaryotic initiation factor 2 (eIF-2α) by activated PKR, and, if provided prior to PKR activation, precluded PKR autophosphorylation. Our study shows that glycosaminoglycans and more specifically heparan sulfate (HS) expressed on the cell surface and extracellular matrix may act as negative regulator of cell-cell fusion.