Termination efficiency of the UAA, UGA and UAG ‘abundant’ and ‘rare’ termination signals in

<p><b>Copyright information:</b></p><p>Taken from "Comparison of characteristics and function of translation termination signals between and within prokaryotic and eukaryotic organisms"</p><p>Nucleic Acids Research 2006;34(7):1959-1973.</p><p>Published online 13 Apr 2006</p><p>PMCID:PMC1435984.</p><p>© The Author 2006. Published by Oxford University Press. All rights reserved</p> Termination efficiency was derived from the 3A′ protein synthesis termination assay (). Readthrough (%) was calculated by comparison of readthrough protein (3A′) to total protein (3A′ + 2A′). Constructs were assayed in specific strains of XAc wild-type strain (dark grey), XA105 UAA suppressor strain (light grey), CDJ64 UGA suppressor strain (light grey) and XA101 UAG suppressor strain (light grey). The mean values from six experiments are presented. Error bars are ±SEM

Termination efficiency of the eukaryotic UAA, UGA and UAG ‘abundant’ and ‘rare’ termination signals and

<p><b>Copyright information:</b></p><p>Taken from "Comparison of characteristics and function of translation termination signals between and within prokaryotic and eukaryotic organisms"</p><p>Nucleic Acids Research 2006;34(7):1959-1973.</p><p>Published online 13 Apr 2006</p><p>PMCID:PMC1435984.</p><p>© The Author 2006. Published by Oxford University Press. All rights reserved</p> Termination efficiency was derived from a dual luciferase protein synthesis termination assay (–) and using COS-7 cells (–). Readthrough (%) was calculated by comparison to a control construct (UGG). The mean values from three experiments are presented. Error bars are ±SEM

Enzymatic inhibition of TcSir2rp1 by BNIP derivatives.

<p><b>A-B)</b> The percentage of inhibition of the NAD⁺-dependent deacetylase activity of TcSir2rp1 by BNIP derivatives is represented in the y axis. Positive (no drug) and negative (NAM, nicotinamide at 2 mM) controls are represented. Bars represent the average + standard deviation of at least two independent experiments. Differences between the experimental groups were considered significant as follows: * p<0.05, *** p<0.001 and **** p<0.0001.</p

Chemical structures of the bisnaphthalimidopropyl derivatives used in the present study.

<p>R = H in all compounds except for compound 13.</p

<i>In vitro</i> activity of BNIP derivatives.

<p><b>A)</b> Primary screening of BNIP derivatives by an <i>in vitro</i> assay for intracellular <i>T</i>. <i>cruzi</i> amastigotes at a single dose of 2.5 μM. Values for both anti-parasitic activity and cell ratio are represented in the dot plot graph. The blue square represents the zone of selection for active, low toxicity hits to be evaluated by dose-response curve analysis. The green square represents the zone of selection for compounds to be tested at a higher dose. <b>B)</b> Primary screening of the compounds selected for testing at a higher dose of 10 μM. Values for both anti-parasitic activity and cell ratio are represented in the dot plot graph. The blue square represents the zone of selection for active, low toxicity hits to be evaluated by dose-response curve analysis. <b>C)</b> Representation of the selectivity indexes of the compounds analysed by dose-response curve analysis, with the determined anti-parasitic activity (EC₅₀) in the y axis, and the cell ratio (CC₅₀) in the x axis. Dots represent the average of three independent experiments.</p

<i>In vitro</i> hepatocytes and neurons injury scores for BNIPSpd (9) by HCS.

<p>The score was calculated as the sum of individual scores obtained from a panel of <i>in vitro</i> cytotoxicity assays that include: mitochondrial dysfunction measured by TMRM probe dynamics in cells; membrane integrity assayed by lactate dehydrogenase quantification; DNA damage by imaging with H2AX antibody; apoptosis by caspase 3/7 activation; and neurite outgrowth as imaged with an anti-tubulin III antibody. Nimesulide (400 μM), an approved drug with a mild toxicological profile, was included as a toxicity control.</p

TcSir2rp1 structural model and docking with compound 9.

<p><b>A</b>) The 3.5 Å structural model shows a large Rossmann-fold domain (composed of 6 parallel β-strands, sandwiched between 2 layers of α-helices), and a small zinc binding domain. The substrate acetylated peptide p53 is bound to the cleft between the small and the large domains. <b>B-C</b>) The substrate p53 peptide was removed from the structure and docking studies conducted with compound <b>9</b>. Several conformations (only 2 shown here) of compound <b>9</b> were possible in a putative ligand binding site close to the NAD binding site.</p

EC₅₀ values (μM) of BNIP derivatives and nifurtimox against <i>T</i>. <i>cruzi</i> epimastigotes, wild-type (WT) and cells overexpressing TcSir2rp1 (OE) with the pTcINDEX.

<p>EC₅₀ values (μM) of BNIP derivatives and nifurtimox against <i>T</i>. <i>cruzi</i> epimastigotes, wild-type (WT) and cells overexpressing TcSir2rp1 (OE) with the pTcINDEX.</p

Anti-parasitic activity, cytotoxicity and selectivity index of BNIP derivatives selected for dose-response curve analysis.

<p>Anti-parasitic activity, cytotoxicity and selectivity index of BNIP derivatives selected for dose-response curve analysis.</p

Cytotoxicity and selectivity of BNIPSpd (9).

<p>Cytotoxicity and selectivity of BNIPSpd (9).</p