Translocation assay

Mouse visible or eleetrophoretie specifie-locus tests Assays for skeletal and cataract mutations Cytogenetic analy.sis and heritable translocation assays DNA damage and repair in rodent germ cells Dominant lethal assay... 

Since the pioneering work of Kleymann et al. (2002), Betz et al. (2002), Baumeister et al. (2007), and Crute et al. (2002), who showed that compounds identified as inhibitors of the helicase-primase enzyme complex could alleviate herpesvirus-induced disease in animal models, the attention of researchers developing antiviral compounds has been drawn more and more towards the virus-encoded helicases, particularly those of Herpes viruses and of RNA viruses such as Hepatitis C Virus (HCV) and SAKS coronavirus (SARS-CoV). Enzyme activity is usually assayed by measuring NTPase activity in the presence of an appropriate nucleic acid co-substrate although, more recently, novel fiuorimetric and luminescence principles have been applied to the measurement of strand unwinding and/or translocation of the protein along the nucleic acid (Frick 2003, 2006). 

Binding to transport proteins may be of particular interest, since binding not only assays the affinities of the binding site on the transporter protein but also the translocation equilibria [67], In terms of enzyme catalysis, a transport protein transforms a substrate, a molecule located at one side of the membrane, into a product, the same molecule at the other side of the membrane, without chemical modification. Substrate must bind to a particular conformation of the enzyme with the binding sites accessible only from, for example, the outside. Similarly, the release of the product has to occur from a conformation which opens the binding site to the inside only this implies at least one transition step between the two types of conformations (see Fig. 

An assay that produces multiple biological readouts. Most commonly used in relation to the mathematical analysis of an image acquired using an automated microscope whereby analysis algorithms quantify cellular parameters (e.g., number, motility, neurite outgrowth, size, shape) and subcellular events (e.g., receptor internalization, protein translocation, protein expression nuclei shape). 

Under basal conditions, PKC is predominantly a cytoplasmic protein. Upon activation by Ca2+ or DAG, the enzyme associates with the plasma membrane, the site of many of its known physiological substrates, including receptors and ion channels. In fact, the translocation of PKC from the cytoplasm to the membrane has long been used as an experimental measure of enzyme activation. Such translocation has often been assayed by phorbol ester binding phorbol esters are tumor-promoting agents that selectively bind to and activate PKC. The molecular basis of the translocation of PKC from the cytoplasm to the plasma membrane has been solved. Subsequent to activation, PKC binds with high affinity to a series of membrane-associated proteins, termed receptors for... 

Mehes G, Pajor L (1995) Nucleolin and fibrillarin expression in stimulated lymphocytes and differentiating HL-60 cells. A flow cytometric assay. Cell Prolif 28 329-336 Nasirudin KM, Ehtesham NZ, Tuteja R, Sopory SK, Tuteja N (2005) The Gly-Arg-rich C-terminal domain of pea nucleolin is a DNA helicase that catalytically translocates in the 5 - to 3 -direction. Arch Biochem Biophys 434 306-315... 

To investigate the effects of drugs on NFkB activation at the molecular level, the Electric Mobility Shift Assay (EMSA) is a useful read-out system. With this technique the nuclear localization of this transcription factor following activation and subsequent translocation can... 

Genetic Toxicology Mouse Heritable Translocation Assay (Original Guideline, adopted 23 October 1986)... 

Genetic toxicology Mouse heritable translocation assay... 

The test detects the ability of a chemical to enhance the exchange of DNA between two sister chromatids of a duplicating chromosome. SCEs are reciprocal interchanges of the two chromatid arms within a single chromosome. These exchanges are visuaUzed during the metaphase portion of the cell cycle and presumably require enzymatic incision, translocation, and ligation of at least two DNA helices. The most commonly used assays employ bone marrow or lymphocytes from mammalian species such as mice, rats, or hamsters. Human lymphocytes may also be used. 

In genotoxic assays both positive and negative results have been reported. A cytogenic study of 35 patients admitted to the hospital after exposure to MIC at Bhopal found no significant effects on sister chromatid exchanges, chromosomal aberrations, or cell cycle. Other studies have found chromosomal abnormalities (especially translocations) in exposed individuals. ... 

Glycidol gave a positive response in assays of prophage induction and SOS repair m Escherichia coli. Results were uniformly positive in several Salmonella typhimurium reverse mutation assays and in two fungal mutation assays. The sex-linked recessive lethal mutation assay and the heritable translocation test in Drosophila melanogaster also gave positive results. 

Lang. R. Adler. I.-D. (1977) Heritable translocation test and dominant-lethal assay in male mice with methyl methanesulfonate. Mutat. Res., 48, 75-88... 

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