It is proposed that the two viral proteins work in concert to promote strand exchange, shown experimentally by generating a gapped circle and a displaced strand from an M13 duplex DNA molecule and an M13 single-stranded circular DNA molecule. However, simian varicella virus (SVV), the primate counterpart of human VZV, does produce chickenpox and reactivate in monkeys. To analyze whether IFNalpha is able to induce such a stably suppressed state of alphaherpesvirus infection, we analyzed whether or not withdrawal of IFNalpha at 5dpi resulted in re-expression of late viral antigens in TG neurons. In agreement with these findings, Solexa/Illumina deep sequencing of small RNAs derived from TG recovered from rhesus macaques latently infected with simian varicella virus (SVV), the primate equivalent of VZV, yielded 1,420,064 usable reads, but, again, did not yield any small RNAs of SVV origin (J. 2K). For immunohistochemical analysis, sections were blocked with Block Ace reagent (DS Pharma Biomedical, Osaka, Japan) and incubated with an anti-HSV-1 antibody (B0114; Dako), which cross-reacts with FBAHV1.
DNA was extracted from each tissue sample with DNAzol and then purified by treatment with phenol-chloroform-isoamyl alcohol (25:24:1). Hence, proteasome activity is required for HSV-2 Us3-mediated PML-NB disruption. High power LEDs centered around 450 nm and 505 nm are readily available, and are a good alternative to arc lamps. These foci may be analogous to HSV-1 VP26 foci, which serve as areas of capsid assembly . Average values are shown. At 16 h after infection, cells were washed with ice-cold PBS and scraped into 500 μl of M-PER (Pierce, Rockford, IL) supplemented with 150 mM NaCl, protease inhibitors (Roche Diagnostics, Germany), and phosphatase inhibitors (20 mM NaF, 10 mM β-glycerophosphate, 2 mM EGTA, and 1 mM Na3VO4).
As a consequence, this high recombination rate allowed us to study the effect of the time interval between infections on the recombination of BoHV-1. The UL sequence is 119,146 bp in length, and the US sequence is 16,405 bp (Table ). The DNA–serum-free medium-Lipofectamine PLUS mix was applied to the cells for 3 h before an equal volume of medium containing twice the normal amount of serum and antibiotics was added. Culture conditions and dissection methods were previously described by Ch’ng et al. We then added DiO, a green fluorescent lipophilic dye, to the N compartment to label all the cell bodies that have extended axons to the N compartment (19). For example, phosphorylation of SG proteins G3BP , growth factor receptor-bound protein 7 (Grb7; ) and dual specificity tyrosine-phosphorylation-regulated kinase 3 (DYRK3; ) all lead to SG disassembly, as does removal of PAR modifications by PAR glycohydrolases .
PRV Ba harbors a deletion removing the entire Us9-coding sequence (35, 42) and was used as a negative control in our experiments. Whereas several tegument proteins like those encoded by HSV-1 open reading frames (ORFs) UL7, UL11, UL16, UL21, UL36, UL37, and UL51 are conserved in all herpesvirus subfamilies, other major tegument components, encoded by UL41, UL46, UL47, UL48, and UL49, are apparently restricted to alphaherpesviruses (54, 56). Using online database annotation tools, we cross-referenced the published gene identifiers to obtain the mouse Entrez Gene ID as a common identifier for each gene (Dennis et al. The first hints that mammalian alphaherpesviruses express miRNAs came from the Tuschl laboratory in 2005 . We also investigated the plasma membrane sites of viral exocytosis, and found no evidence for specialized cytoskeleton-depleted exocytosis sites, contrary to a previous report on HSV-1 . Dissociated rat superior cervical ganglion (SCG) neurons were cultured in vitro under tissue culture conditions where only axons, but not dendrites, are produced (Ch’ng et al., 2005; Tomishima and Enquist, 2001).
Synthetic siRNA’s have been shown to successfully inhibit viral replication of several viruses, among them herpesviruses –. These proteins localize to the outer mitochondrial membrane and have two calcium-binding EF hand domains that regulate the activity of both kinesin-1 and dynein, as well as the interactions of these motors with mitochondrial protein complexes (Fransson et al., 2006). However, VZV ORF66 has a substrate specificity similar to that of HSV-1 and PRV US3 kinases in that it preferentially targets S/T residues embedded in basophilic motifs. Specific PRV gene products have been identified as virulence factors and these research efforts have led to successful live vaccines, such as the strain PRV Bartha and its derivatives (see ref.