[Expression of herpes simplex virus type 2 latency associated transcript ORF1 and its anti-apoptotic function]
Summary
Herpes simplex virus type 2 (HSV-2) latency-associated transcript open reading frame 1 (LAT ORF1) is expressed in Vero cells. This gene protects cells from actinomycin D-induced apoptosis.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- Herpes simplex virus type 2 (HSV-2) establishes latency in neurons.
- Latency-associated transcript (LAT) is crucial for HSV latency.
- The function of LAT open reading frame 1 (ORF1) in apoptosis is not fully understood.
Purpose of the Study:
- To investigate the expression of HSV-2 LAT ORF1 in Vero cells.
- To determine the anti-apoptotic function of HSV-2 LAT ORF1 when induced by actinomycin D.
Main Methods:
- Construction and transfection of recombinant plasmid pEGFP-ORF1 into Vero cells.
- RT-PCR for gene expression analysis.
- Fluorescence microscopy and Hochest33258 staining for cell morphology.
- MTT assay for cell viability.
- Flow cytometry for apoptosis rate detection.
Main Results:
- Successful construction and high expression of pEGFP-ORF1 in Vero cells confirmed by RT-PCR.
- Vero cells transfected with pEGFP-ORF1 showed no morphological changes and maintained viability comparable to controls when treated with actinomycin D.
- Apoptosis rates were significantly lower in cells expressing pEGFP-ORF1 compared to empty vector controls.
Conclusions:
- HSV-2 LAT ORF1 gene is expressed in Vero cells.
- HSV-2 LAT ORF1 confers protection against actinomycin D-induced apoptosis in Vero cells.
Related Concept Videos
01:17The Extrinsic Apoptotic Pathway
6.2K
The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...
6.2K
02:28Leaky Scanning
4.5K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R...
4.5K
01:30Inhibitors of Viral Protein Synthesis
58
Protein synthesis is indispensable for viral replication, as viruses lack the cellular machinery required for this process and must hijack the host's translational apparatus. In response, host cells deploy a critical innate immune defense involving interferons, specialized cytokines that play a central role in inhibiting viral propagation.Upon viral detection, infected cells release interferons that bind to receptors on adjacent uninfected cells, activating the JAK-STAT signaling pathway and...
58
01:31The Intrinsic Apoptotic Pathway
6.2K
Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...
6.2K
01:30Apoptosis
12.0K
Apoptosis is a combination of two Greek words, 'apo' and 'ptosis,' meaning separation and falling off, respectively. Hippocrates used this word to describe gangrene, which was caused due to bandaging of fractured bones. Apoptosis was distinguished from necrosis in 1970 when John Kerr reported observations of morphological changes occurring during apoptosis. During one experiment, he observed that the disruption of blood supply to the liver tissue resulted in a size...
12.0K