2026-10-14T13:35:44Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34035570 2026-09-30T17:35:04Z category_4 category_12 category_13 category_14 category_734 portal_5 item_type_1 month_year_09_2026

<p>Screen for suppressors of the <i>orc4-R267A</i> lethal phenotype.</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) An <i>orc4-R267A</i> strain kept alive by a plasmid containing <i>ORC4</i> was treated with ethyl methane sulfonate (EMS) to isolate suppressors of the lethal phenotype of the <i>orc4-R267A</i> allele. Only strains that acquire suppressors can lose the plasmid. The desired strains turn red and grow on 5-Fluorouracil (5-FOA) due to the loss of the <i>ADE2</i> and <i>URA3</i> genes on the plasmid. B) Table of <i>orc4-R267A</i> suppressor mutations identified in the screen. C) Ten-fold serial dilutions of strains that have <i>ORC4</i> on a counter-selectable plasmid and either <i>ORC4</i> or <i>orc4-R267A</i> allele integrated into the genome. The presence of suppressors-of-orc-four-ATPase alleles are indicated. Some strains are red prior to 5-FOA selection because the suppressor allele allowed plasmid loss during strain construction.</p> 2026-09-30T17:34:54Z Image Figure 10.1371/journal.pone.0349465.g001 https://figshare.com/articles/figure/_p_Screen_for_suppressors_of_the_i_orc4-R267A_i_lethal_phenotype_p_/34035570 CC BY 4.0