Herpes simplex virus type 1 replication and recombination. – PubMed

We now demonstrate that replication compartments can be formed by cotransfecting Vero cells with constructs expressing the seven essential viral replication proteins and a plasmid containing an HSV-1 origin of DNA replication. DAI colocalized with ICP0 in a subset of nuclear and cytoplasmic foci in infected cells and interacted with ICP0 in coimmunoprecipitation assays. This report demonstrates that cellular responses to chemotherapeutic agents provide an unfavorable environment for HSV-1-mediated oncolysis, and these observations are relevant to the design of both preclinical and clinical studies of HSV-1 oncolysis. Thus, both MLH1 and MSH2 are required but appear to participate in distinct events in the virus life cycle. contributed equally to the work reported in this article. Similarly, the promoter for the DF3/MUC1 tumor-associated antigen was cloned into a third HSV-1 mutant such that it regulates expression of gamma(1) 34.5. Virol.

These data indicate that several key events in which ICP27 plays a role are curtailed during infection with ICP27 phosphorylation site mutants. The images were obtained by confocal microscopy. Activation of NF-κB usually involves proteasome-dependent degradation of IκBα after it has been phosphorylated by the IκB kinase (IKK). These data indicate that several key events in which ICP27 plays a role are curtailed during infection with ICP27 phosphorylation site mutants. This result suggests that C11AG interferes with cellular signal transduction mechanisms that regulate expression of HSV-1 immediate early genes. The HSV expression cascade during replication consists of the expression of (1) immediate early genes (IE) and (2) early (E) genes from the circularized HSV genome, of (3) leaky late genes (LL) from the parental genome and progeny genomes and of (4) late genes (L) from progeny genomes. Lower, HSV-1 replication causes a series of structural changes in the cell nucleus characterized by co-localization of replication, repair, and recombination proteins (for further details, see Ref.


DAI colocalized with ICP0 in a subset of nuclear and cytoplasmic foci in infected cells and interacted with ICP0 in coimmunoprecipitation assays. Everett, Virology 217:67-75, 1996). Cell-specific DNA synthesis was also adversely affected, but this did not alter cell viability or plating efficiency. Notably, ICP4-containing replication compartment formation was severely compromised, with the appearance of small ring-like structures that persisted even at late times after infection. Although ICP0 has been shown to be important for partial inactivation of other cellular DNA repair pathways, we show that ICP0 is not responsible for the inactivation of ATR signaling and, furthermore, that neither ATR nor ATRIP is a target of ICP0 degradation. These data show that while ICP34.5 regulates autophagy, it is the prevention of translational arrest by ICP34.5 rather than its control of autophagy that is the pivotal determinant of efficient HSV-1 replication in primary cell culture. Together, these data suggest that ATR pathway proteins are not antiviral per se but that activation of ATR signaling may have negative consequences during viral replication, such as inhibiting recombination.

Therefore, the effect of viral infection on the formation of paraspeckles is probably attributable to the increase in NEAT1 expression, which facilitates the organization of the paraspeckle components. (C and D) Vero cells stably expressing either an empty vector, Flag-ATR, HA-ATRIP, or HA-ATRIP-CRD were generated by retroviral infection and continuous selection in puromycin. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. of white blood cells, adrenalin etc. However, the extent to which ICP34.5-deficient HSV-1 replicates in and may be neurotoxic to normal brain cell types in vivo is poorly understood. This book presents chapters on protocols on many strands of HSV-1 research that are currently in use in leading laboratories. (B) Vero cells stably expressing Myc-HCLK2 were generated by retroviral infection and continuous selection in puromycin.

When this cellular response is abrogated, formation of HSV-1 replication centers is retarded, and viral production is compromised. The product of one of the essential seven genes, the UL 9 protein, serves as the replication initiator, which binds to and unwinds DNA at a viral origin. Much of the infectious cycle occurs in the nucleus of cells where the virus has evolved mechanisms to manipulate host processes for the production of virus. We have constructed recombinant baculoviruses which efficiently express the products of each of these seven genes in infected Spodoptera frugiperda (Sf) insect cells. From a bioassay-guided fractionation procedure, PS-A-6 was isolated from Psychotria serpens (P. DNA damage-induced ATR activation slightly reduces HSV-1 replication. tsBN2, a temperature-sensitive (ts) growth mutant of the hamster cell line BHK-21, has a point mutation in the RCC1 (regulator of chromosome condensation) gene, and prematurely enters mitosis at 39.5 degrees C, a nonpermissive temperature.

Effects of Sam B on HSV-1 replication and Vero cell viability and growth. Vero cells and L7 cells [52] were propagated in Dulbecco’s Modified Eagle’s medium (DMEM) containing 0.15% HCO3- supplemented with 5% fetal bovine serum (FBS), penicillin G (100 U/ml), and streptomycin (100 mg/ml), hereafter referred to as “complete DMEM.” Wild-type HSV-1 strains KOS, KOS-GFP, and McKrae were propagated in Vero cells cultured in complete DMEM.