Unfortunately, HIV-1 infections cannot be cured, so that drug therapy, once initiated, must be continued for the life of the patient. Mapping of the regions involved in binding and transcriptional activation showed that the amino terminus of LANA is the major site for interaction and up-regulation but that it can cooperate with the carboxy terminus to enhance these functions. Get a printable copy (PDF file) of the complete article (965K), or click on a page image below to browse page by page. Poly(A)+ RNA was treated with the Access RT-PCR kit (Promega, Madison, Wis.). Electrophoretic mobility shift assays indicated that the latency-associated nuclear antigen targets and affects the Sp1-DNA complex in the context of BJAB nuclear extracts. Our study provides the first evidence that expression of TR, in this case encoded by a herpesvirus, is pro-oncogenic in the absence of telomerase activity.
For many years there existed a paradigm in molecular biology known as the “central dogma.” This asserted that DNA is first transcribed into RNA, RNA is translated into amino acids, and amino acids assemble into long chains, called polypeptides, that make up proteins—the functional units of cellular life. Natl. Mapping of the regions involved in binding and transcriptional activation showed that the amino terminus of LANA is the major site for interaction and up-regulation but that it can cooperate with the carboxy terminus to enhance these functions. HHV-6A(GS) strain was propagated in the human T-cell line HSB-2; HHV-6B(PL1) strain was propagated in activated peripheral blood mononuclear cells (PBMC). Despite the existence of an effective vaccine, there is no completely effective antiviral treatment for patients chronically infected with hepatitis B virus (HBV). This study documents that the RT-PCR assay can successfully identify LATs and may serve as a tool to better understand the biologic characteristics of FHV-1 and its relationship to clinical disease.
In conclusion, this class of indolopyridones can occupy the nucleotide binding site of HIV RT by forming a stable ternary complex whose stability is mainly dependent on the nature of the primer 3′ end. Electrophoretic mobility shift assays indicated that the latency-associated nuclear antigen targets and affects the Sp1-DNA complex in the context of BJAB nuclear extracts. It also had different impacts on the actions of the L-nucleoside triphosphates toward DPs. The chameleon-like nature of HIV, however, limits their continued effectiveness. However, this system does not permit analysis of HBV mutants that are now being reported, because these cells are stably transfected with wild-type HBV. Schleif, K.
Our findings suggest that physiological pathways that include these cellular factors may be involved in modulating HSV reactivation. Human herpesvirus (HHV) infections are commonly associated with HIV-1. Five adults ranged from 40–81 years of age. Pretreatment of HLFs with deoxynucleosides, which increase cellular dNTP pools, enabled the mutant vector to transduce HLFs, suggesting that the transduction failure of the RT mutant vector to primary cells is because of inefficient reverse transcription in low cellular dNTP environments. The limit of detection (LOD95% ) calculated by probit regression analysis was 23 copies per reaction. BARF1 expression was found in 6 of 7 nasopharyngeal carcinomas (NPC) but in 0 of 22 Hodgkin’s disease (HD) cases, whereas LMP2 expression was found in 7 of 7 NPCs and in 17 of 22 HD cases.
Finally, to assess infectious viral production, other primary human cells (human umbilical vein endothelial cells), were infected with concentrated supernatant of KSHV-infected, TPA-induced keratinocytes and the presence of viral transcripts was confirmed by RT-PCR. After baseline evaluation, 19 patients received AZT, 23 AZT plus DDI, 3 started AZT only with DDI added after 3 months, and 3 received a combination of AZT plus 3TC. An RT-PCR assay detecting 4 different HHV-6 gene transcripts was established. The viral latency program of KSHV is central to persistent infection and plays important roles in the pathogenesis of KSHV-related tumors. We identified and validated an interaction between HIV-1 RT and the RNA-binding protein HuR. For further characterization of virus reactivation and monitoring of viral transcription we established real-time RT-PCR assays using TaqMan technology to sensitively quantify viral transcripts expressed at different times of the lytic cycle: for BZLF1, an immediate early transactivator initiating the transition from latency to lytic replication, for the DNA-polymerase BALF5 and for the major viral glycoprotein gp350/220 (BLLF1).