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oai:figshare.com:article/34035576 2026-09-30T17:35:06Z category_4 category_12 category_13 category_14 category_734 portal_5 item_type_1 month_year_09_2026

<p>Biochemical effects of suppressor mutations on the biochemical activities of the ORC4R protein.</p> Luis E. Martínez-Rodríguez (25145346) Stephen P. Bell (25145349) Biochemistry Cell Biology Genetics Molecular Biology Biological Sciences not elsewhere classified origin recognition complex saccharomyces cerevisiae </ helicase loading activity div >< p atpase interfaces within six orc subunits orc atp hydrolysis defective orc allele atp hydrolysis orc subunits defective orc vitro </ toa2 </ r267a </ sixth allele reducing transcription orc4r ). lethal phenotype genetic screen essential role cdc6 complex binds origins although functions <p>A) ATPase activity of WT ORC and ORC4R protein with or without the indicated <i>sofa</i> mutation. Hydrolysis was monitored using radioactive ATP. Percent hydrolysis (ADP/ (ADP + ATP), arbitrary units) was determined over a time course. Plots display the mean and standard deviation of three replicates as points and errors bars, respectively. B) Electrophoretic-mobility shift assay to determine <i>ARS1</i> DNA binding of mutant protein complexes. Representative experiments are shown for each experimental set. Bound fraction was normalized within each gel to the WT + ATP reference and plotted for three replicates of each experimental set. WT data is displayed twice. C) <i>In vitro</i> helicase-loading assay for WT and ORC4R protein without or with the indicated suppressor mutation. Potassium glutamate concentration was 350mM. Representative experiment is shown. Percent of WT was quantified for three replicates.</p> 2026-09-30T17:34:54Z Image Figure 10.1371/journal.pone.0349465.g003 https://figshare.com/articles/figure/_p_Biochemical_effects_of_suppressor_mutations_on_the_biochemical_activities_of_the_ORC4R_protein_p_/34035576 CC BY 4.0