Nuclear Protein Kinases
Lubomir S. Hnilica in Chromosomal Nonhistone Proteins, 2018
The eukaryotic RNA polymerases are another class of proteins involved in gene transcription that are subject to phosphorylation. In yeast five subunits of polymerase I, two subunits of polymerase II, and three subunits of polymerase III are labeled after incubation of cells with 32P-orthophosphate.10,11,177 All three polymerases share a common subunit of 24,000 daltons which is phosphorylated. Phosphorylation of RNA polymerase has also been clearly established in vitro in yeast,10,11 rat liver,178,179 and calf thymus,180,181 though in the case of yeast it is interesting to note that in addition to the polymerase subunits normally phosphorylated in vivo, an extra subunit of polymerase I and one from polymerase II are also phosphorylated in vitro.
The Primer Hypothesis for the Regulation of Eukaryotic Gene Expression
M. Gerald, M.D. Kolodny in Eukaryotic Gene Regulation, 2018
Studies71 of eukaryotic RNA polymerase have also shown only a low level of RNA synthesis with native DNA as template because of a failure to initiate RNA chains. It may be that activator RNA, absent in these in vitro studies and acting as the initial sequences in the RNA to be transcribed, provides the necessary initiation for the RNA chains. Investigations of eukaryotic polymerase referred to in a previous section have shown that RNA synthesis is not initiated on synthetic polynucleotide templates.72 Perhaps activator RNA associated with extracted DNA is necessary to initiate transcription. Kanehisa et al.73-74 have shown that chromatin low molecular weight RNA from chick liver had specific stimulatory effects on RNA polymerase when chick liver chromatin was used as template but inhibitory activity when E. coli DNA was used as template. Perhaps activator RNA specific for chick was present amongst the chromatin RNA.
Methods in molecular exercise physiology
Adam P. Sharples, James P. Morton, Henning Wackerhage in Molecular Exercise Physiology, 2022
Inherited genetic variations or epigenetic modifications, as a result of exercise, are able to effect whether a gene is turned on or off. The process of turning a gene on or off is termed gene transcription or expression (the full theory for gene transcription is described in detail in Chapter 3). In order to measure gene expression, RNA must first be isolated. As with DNA, researchers are able to directly extract RNA from skeletal muscle tissue to study gene expression following exercise. RNA is very similar to DNA; however, RNA is single-stranded, whereas DNA is double-stranded (i.e. the double helix structure); the sugar backbone in RNA is ribose compared with deoxyribose within DNA (hence why RNA is much less stable than DNA) and there is a replacement of thymine (T) in DNA with uracil (U) in RNA (see Chapter 3, Figure 3.7). Also known as mRNA, the RNA copies and delivers the DNA ‘message’ to the protein-making machinery of the cell (i.e. the ribosome) to initiate the production of protein. Therefore, as suggested above, the making or synthesis of RNA from DNA is termed transcription, whereas the synthesis of protein from transcribed mRNA is termed translation.
Novel immunotherapeutic approaches for hepatocellular carcinoma treatment
Published in Expert Review of Clinical Pharmacology, 2019
Davide Busato, Monica Mossenta, Lorena Baboci, Federica Di Cintio, Giuseppe Toffoli, Michele Dal Bo
RNA production is a rapid, not expensive and validated procedure that enables messenger RNA synthesis encoding for many types of transcripts. This characteristic can be useful to setup vaccination strategies due to the possibility of including more tumor neo-epitopes to elicit an optimal immune response [205,206]. Moreover, RNA is rapidly degraded decreasing both side effects and autoimmune diseases [204]. A mRNA-based vaccine is ongoing in a phase I/II clinical trial to treat several types of cancer including HCC (NCT03480152). It provides the injection of NCI-4650 vaccine, a personalized vaccine presenting up to 15 neoantigens specific of patient malignancy [207]. Peptide-based vaccines have the advantage to be cost-effective and to use pre-defined epitopes useful to obtain the best patient immune response. In contrast, the TAAs that can be recognized are few with the possibility to have low antigen-specific activation of CD8+ T cells and/or CD4+ T cells [208]. This leads to a low vaccine efficacy [206,208].
Vitexin protects melanocytes from oxidative stress via activating MAPK-Nrf2/ARE pathway
Published in Immunopharmacology and Immunotoxicology, 2020
Xiao-Sha Li, Xue-Yong Tang, Wei Su, Xin Li
Total RNA was isolated from melanocytes using a total RNA extraction kit (Anmei Biologicals Ltd, Xi’an, China), following the manufacturer’s instructions. The concentration and purity of isolated RNA were determined by measuring the absorbance at an optical density of 260 nm by using a spectrophotometer (Bio-Rad, Hercules, CA). And then isolated RNA served as a template for cDNA synthesis using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Waltham, MA), following the manufacturer’s instructions. qPCR was performed with SYBR Premix Ex Taq II (TaKaRa, Ohtsu, Japan) per vendor’s instructions to determine the relative mRNA levels of various target genes. Briefly, the cycling conditions were set as start at 95 °C for 2 min, then 43 cycles of amplification. Each cycle includes denaturation (95 °C for 5 s), annealing (60 °C for 10 s), and extension (72 °C for 15 s). All reactions were performed in triplicate and analyzed using the 2−ΔΔCt method. GAPDH was used as an internal control. The following primer sets were designed and synthesized by GenScript (NanJing, China):
Evaluation of the toxicological effects of favipiravir (T-705) on liver and kidney in rats: biochemical and histopathological approach
Published in Drug and Chemical Toxicology, 2023
Adem Kara, Seda Yakut, Cuneyt Caglayan, Tuğçe Atçalı, Aykut Ulucan, Fatih Mehmet Kandemir
Favipiravir is an RNA polymerase (RNAP) inhibitor which is a family of enzymes and responsible for transcribing the information in a DNA or RNA molecule as an RNA molecule. The process of copying the information contained in a gene as an RNA molecule is called transcription. RNAP enzymes found in many viruses and living things allow genes to be read as RNA strands in the cells. Molecular studies revealed that viral proteases play a critical role in the life cycle of many viruses, either by influencing the cleavage of high molecular weight viral polyprotein precursors for the functional products or by catalyzing the processing of structural proteins. Nowadays, many types of research are conducted on protease inhibitors to treat infections caused by many RNA and DNA viruses, such as HIV, HCV, Picornaviruses, RSV, Herpes viruses, Rotavirus, and SARS (Mendenhall et al. 2011b, Goldhill et al. 2018, Musa 2020).
Related Knowledge Centers
- Nucleic Acid
- Nucleotide
- Polymerase
- Primary Transcript
- Protein
- Messenger Rna
- NON-Coding Rna
- Base Pair
- Complementarity
- Antiparallel