133,027 research outputs found
MUT-16 and orthologs contain a high degree of predicted disorder.
No full text(A) Graph comparing disorder tendency of C. elegans MUT-16 (isoform b, WP:CE40347) in red to orthologs in C. remanei (RP:RP48608) in light gray, C. briggsae (BP:CBP44329) in medium gray, and C. japonica (JA:JA63728) in dark gray using IUPRED and long disorder parameters (http://iupred.enzim.hu/). Scores above 0.5 indicate disorder. Residue positions correspond to the C. elegans MUT-16 sequence, but all proteins are similar in length. (B) Q, N, and P residues in C. elegans MUT-16 were counted in amino acid 100-mers, starting at position one, shifting 10 residues at a time, and displayed as stacked columns. Indicated residue positions are the mid-point of the 100-mer. (C) Schematic of known dependencies of Mutator foci formation. The requirements for localization of RRF-1, RDE-8, NYN-1, NYN-2, and SMUT-1 were not known prior to this work. (D, E) Diagram (D) and table (E) of MUT-16 deletions generated by CRISPR. Bars in (D) are drawn to scale relative to residue positions in (A,B).</p
Histogram of D<sub>mut</sub> for experimental datasets.
No full text<p>D<sub>mut</sub> quantifies expression differences between orthologs (interspecies sets) or the same gene in various conditions (intraspecies sets). Interspecies analysis was performed using wine yeast strains (red) and <i>Drosophila</i> species (green), and intraspecies analysis for <i>Arabidopsis thaliana</i> (blue) and human (purple). The observed 1th and 99th percentile were used to obtain cutoffs for D<sub>mut</sub> in order to describe ‘small’ and ‘large’ changes (D<sub>small</sub> and D<sub>large</sub>).</p
Simulated competition between the wild-type, MUT-H275Y and MUT-I223V strains.
No full text<p>The infectious (PFU/mL) and total (RNA/mL) viral load (top), and the fraction of infectious cells infected by each strain (bottom) are shown. (A) Without therapy, the fitness of the WT and MUT-H275Y strains is comparable. (B) In the presence of oseltamivir, the WT has a replicative disadvantage over the MUT-H275Y. (C) Without therapy, the MUT-H275Y strain already has a fitness advantage over the MUT-I223V strain. (D) In the presence of oseltamivir, this advantage is increased.</p
Facsímil: Insula Maioricae / D. Vicentius Mut deli.
No full text1 Mapa, b/n. - (Col. Alomar; 1). - Reimpreso en Madrid por Adolfo Rupérez, de la Calcografia Nacional, a expensas de la Excma. Diputación de Baleares, en 1946. - Planxa 42 x 58 cm.[1:123 000 aprox]44 x 60 c
Facsímil: Insula Maioricae / D. Vicentius Mut deli.
No full text1 Mapa, b/n. - (Col. Alomar; 1). - Reimpreso en Madrid por Adolfo Rupérez, de la Calcografia Nacional, a expensas de la Excma. Diputación de Baleares, en 1946. - Planxa 42 x 58 cm.[1:123 000 aprox]44 x 60 c
Temperature-dependent dynamic deactivation of pol V Mut wt.
No full textActivity and dynamic deactivation of pol V Mut wt (200 nM) were measured as shown in Fig 1A at (A) 37°C and (B) 30°C on 12 nt oh HP (1 μM) in the presence of saturating concentration of ATPγS (500 μM) and dNTP’s (mix of dTTP, dCTP, dGTP 500 μM each). (C) Prior to deactivation, pol V Mut wt performs one round of DNA synthesis at 37°C (black circles in the graph) and 3 rounds at 30°C (white triangles in the graph). Deactivated pol V Mut is reactivated by adding RecA* wt (200 nM). (D) Activity of pol V Mut wt measured in the presence of ATP (500 μM) and dNTP’s (dTTP, dCTP, dGTP 500 μM each). Pol V Mut wt is not active with ATP but it can synthesize DNA when trans-activated by RecA* wt (trans-activation reaction was performed for 30 min). Representative DNA synthesis gels for pol V Mut wt are presented in (A, B, and D). The experiments were repeated 3 times and the average % PE (percent p/t DNA extended) with the SD (standard deviation) for each reaction time point is graphed in panel C.</p
<i>Taco1</i><sup><i>mut/mut</i></sup> mice generate a normal immune response.
No full text(A) Body weights from uninfected and MCMV infected Taco1wt/wt and Taco1 mut/mut mice. Spleens and livers from uninfected and MCMV infected 30 week old Taco1wt/wt and Taco1 mut/mut mice were prepared for flow cytometry to determine (B) the number of total spleen lymphocytes, (C) the number of spleen T cells, (D) the number of spleen NK cells, (E) the number of total liver lymphocytes, (F) the number of liver T cells, (G) the number of liver NK cells, and (H) the number of m38+ CD8+ T cells in the spleen and liver. The data are representative of results obtained from at least 5 mice from each genotype and each infection group. Error bars indicate SEM; *pt-test.</p
Static deactivation of pol V Mut wt at 37°C and 30°C.
No full textStatic deactivation of pol V Mut wt was measured as shown in Fig 4A. Pol V Mut wt deactivates statically in the absence of DNA synthesis at 37°C (A-B) and 30°C (C-D). Pol V Mut wt was incubated either alone (I. and black circles in the graphs) or with ATPγS (II. and white circles in the graphs), or with ATPγS and 12nt oh HP DNA (III. and black triangles in the graphs). The static deactivation conditions and analysis are described in the legend for Fig 4. Pol V Mut wt deactivates more slowly at 30°C (C-D) compared to 37°C (A-B). However, in contrast to pol V Mut E38K/ΔC17 (Fig 4), the degree of stabilization with ATPγS is much less for pol V Mut wt. Deactivated pol V Mut wt is reactivated by incubation with RecA* (+RecA*).</p
Deactivated pol V Mut E38K/ΔC17 does not bind to p/t DNA.
No full text(A) Pol V Mut was assembled using AF647-labeled RecA E38K/ΔC17. To deactivate pol V MutAF647E38K/ΔC17, 1 μM of enzyme was incubated either alone (lane 2), or with ATPγS (500 μM) (lane 4), or with ATPγS (500 μM) and p/t DNA (5 μM) (lane 6) for 4 hr at 37 oC. After incubation, the activity of pol V Mut E38K/ΔC17AF647 was measured on 32P-labeled 12 nt oh HP DNA in the presence of ATPγS (A), and binding to p/t DNA was detected in smFRET experiments (B-D). Statically deactivated pol V Mut E38K/ΔC17AF647 cannot incorporate dNTPs after 4 hr incubation at 37°C (A, lane 2). ATPγS stabilizes pol V Mut E38K/ΔC17AF647 and protects it from deactivating (A, lane 4). However, deactivation does occur following the addition of DNA to ATPγS-bound pol V MutAF647E38K/ΔC17 (A, panel 6). Deactivated pol V Mut E38K/ΔC17AF647 can be reactivated by RecA* (lanes 3, 5, 7). Pol V Mut E38K/ΔC17AF647 that has not been subject to incubation at 37°C (4 hr incubation on ice) is able to synthesize DNA in the presence of ATPγS (lane 1). (B-D) Representative smFRET images depicting binding of deactivated pol V Mut E38K/ΔC17AF647 to p/t DNA. AF555-labeled p/t DNA is shown in green, pol V Mut E38K/ΔC17AF647 is shown in red. Binding events are shown as colocalized pol V Mut E38K/ΔC17AF647 and p/t DNA yellow/orange signals and are marked with arrows. Representative smFRET trajectories are shown below each deactivation panel (B-D). No DNA binding events are detected for deactivated pol V Mut E38K/ΔC17AF647 (B), which is consistent with the absence of polymerase activity (A, lane 2). Pol V Mut E38K/ΔC17AF647 activity is stabilized substantially in the presence of ATPγS, which results in DNA binding (C) and polymerase activity (A, lane 4). Fewer binding events are detected when pol V Mut E38K/ΔC17AF647 is incubated in the presence of ATPγS + p/t DNA (D) compared to incubation with ATPγS in the absence of DNA (B, middle panel). For all smFRET binding assays, ATPγS (500 μM) was added prior to injection of labeled pol V Mut. The images shown in (B-D) panels correspond to smFRET data integrated over 1 min. Scale bar is 150 mm.</p
Static Deactivation of pol V Mut E38K/ΔC17 at 37°C and 30°C.
No full text(A) Sketch showing static deactivation of pol V Mut E38K/ΔC17 (200 nM) in the absence of DNA synthesis at 37°C and 30°C. Static deactivation was determined by measuring the extent of DNA synthesis as a function of incubation time on 12 nt oh HP p/t DNA at 37°C in the presence of ATPγS and dNTP’s (dTTP, dCTP, dGTP 500 μM each). Pol V Mut E38K/ΔC17 was incubated either alone (I.) or with ATPγS (II.), or with ATPγS and 12nt oh HP DNA (III.). (B) and (D) show representative gels for each deactivation condition. Pol V Mut E38K/ΔC17 deactivates more slowly at 30°C (D-E) compared to 37°C (B-C). At each temperature, pol V Mut is stabilized when incubated in the presence of ATPγS. Deactivated pol V Mut E38K/ΔC17 is not dead since it can be reactivated by incubation with RecA* (+RecA*). Static deactivation is expressed as the relative polymerase activity measured at each incubation time point divided by the polymerase activity measured at t = 0. Each experiment was repeated 2–3 times and average relative activity along with SD for each deactivation time point are presented in (C) and (D). (I. and black circles in the graphs) represent static deactivation of pol V Mut alone, (II. and white circles in the graphs) represent deactivation of pol V Mut +ATPγS and (III. and black triangles in the graphs) represent deactivation of pol V Mut + ATPγS + 12 nt oh HP.</p
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