RPA inhibits the decatenase activity of both TopoIIIα and EcTop1.

<p>(<b>A</b>) Decatenation reactions containing TopoIIIα (30 nM, lanes 2–4) and RPA (100 nM, lane 3; 200 nM, lanes 4 and 5) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (TopoIIIα alone). The percent of catenated substrate converted to circular products is indicated. (<b>B</b>) Decatenation reactions containing EcTop1 (6 nM, lanes 2–4) and RPA (100 nM, lane 3; 200 nM, lanes 4 and 5) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (EcTop1 alone). The percent of catenated substrate converted to circular products is indicated.</p

TopoIIIα, BLM and RMI1 cooperate to catalyze decatenation on RPA-coated single-stranded catenane.

<p>(<b>A</b>) In a competitive binding model, TopoIIIα alone is unable to access RPA-coated substrate. (<b>B</b>) Complex formation by BLM, TopoIIIα, and RMI1 enables displacement of RPA from the substrate. (<b>C</b>) Alternatively, the DNA binding activity of BLM drives the complex to the substrate even when the substrate is coated with RPA, promoting decatenation.</p

BLM alleviates EcSSB inhibition of TopoIIIα, but not EcTop1, decatenase activity.

<p>(<b>A</b>) Decatenation reactions containing TopoIIIα (30 nM, lanes 2–8), EcSSB (3.2 mM, lanes 3–9), BLM (33 nM, lanes 4, 8 and 9; 66 nM, lane 5) and RMI1 (100 nM, lanes 6, 8 and 9; 200 nM, lane 7) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (TopoIIIα alone). The percent of catenated substrate converted to circular products is indicated. (<b>B</b>) Decatenation reactions containing TopoIIIα (20 nM, lanes 2–9), RPA (200 nM, lanes 3–5 and 10), EcSSB (3.2 mM, lanes 6–8 and 10) and BLM (33 nM, lanes 4 and 7; 66 nM, lanes 5 and 8–10) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (TopoIIIα alone). The percent of catenated substrate converted to circular products is indicated. (<b>C</b>) Decatenation reactions containing EcTop1 (6 nM, lanes 2–7), EcSSB (100 nM, lanes 3–6 and 8), BLM (17 nM, lanes 4–8) and RMI1 (75 nM, lane 5; 150 nM, lanes 6–8) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (EcTop1 alone). The percent of catenated substrate converted to circular products is indicated.</p

BLM-RMI1 alleviates RPA inhibition of TopoIIIα decatenase activity.

<p>(<b>A</b>) Decatenation reactions containing TopoIIIα (30 nM, lanes 2–7), RPA (100 nM, lanes 3–8), BLM (33 nM, lane 4; 66 nM, lanes 5 and 9) and RMI1 (100 nM, lane 6; 200 nM, lanes 7 and 10) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (TopoIIIα alone). The percent of catenated substrate converted to circular products is indicated. (<b>B</b>) Decatenation reactions containing TopoIIIα (15 nM, lanes 2–8), RPA (100 nM, lanes 3–6, 8 and 9), BLM (17 nM, lanes 4–9), wild type RMI1 (75 nM, lane 5; 150 nM, lanes 6, 7 and 9) and RMI1-LLTD mutant (150 nM, lanes 8 and 10) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (TopoIIIα alone). The percent of catenated substrate converted to circular products is indicated. (<b>C</b>) Decatenation reactions containing TopoIIIα (7.5 nM, lanes 2–4), BLM (8 nM, lanes 2–4), RMI1 (38 nM, lanes 2–4) and RPA (140 nM, lane 3; 280 nM, lane 4) as indicated were fractionated on a denaturing polyacrylamide gel and autoradiographed. Quantification of the decatenation products is presented in the histogram, normalized to the reactions in lane 2 (TopoIIIα-BLM-RMI1). The percent of catenated substrate converted to circular products is indicated.</p

Comparative Chemogenomics To Examine the Mechanism of Action of DNA-Targeted Platinum-Acridine Anticancer Agents

Platinum-based drugs have been used to successfully treat diverse cancers for several decades. Cisplatin, the original compound of this class, cross-links DNA, resulting in cell cycle arrest and cell death <i>via</i> apoptosis. Cisplatin is effective against several tumor types, yet it exhibits toxic side effects and tumors often develop resistance. To mitigate these liabilities while maintaining potency, we generated a library of non-classical platinum-acridine hybrid agents and assessed their mechanisms of action using a validated genome-wide screening approach in <i>Saccharomyces cerevisiae</i> and in the distantly related yeast <i>Schizosaccharomyces pombe.</i> Chemogenomic profiles from both <i>S. cerevisiae</i> and <i>S. pombe</i> demonstrate that several of the platinum-acridines damage DNA differently than cisplatin based on their requirement for distinct modules of DNA repair

Comparative Chemogenomics To Examine the Mechanism of Action of DNA-Targeted Platinum-Acridine Anticancer Agents

Platinum-based drugs have been used to successfully treat diverse cancers for several decades. Cisplatin, the original compound of this class, cross-links DNA, resulting in cell cycle arrest and cell death <i>via</i> apoptosis. Cisplatin is effective against several tumor types, yet it exhibits toxic side effects and tumors often develop resistance. To mitigate these liabilities while maintaining potency, we generated a library of non-classical platinum-acridine hybrid agents and assessed their mechanisms of action using a validated genome-wide screening approach in <i>Saccharomyces cerevisiae</i> and in the distantly related yeast <i>Schizosaccharomyces pombe.</i> Chemogenomic profiles from both <i>S. cerevisiae</i> and <i>S. pombe</i> demonstrate that several of the platinum-acridines damage DNA differently than cisplatin based on their requirement for distinct modules of DNA repair

Comparative Chemogenomics To Examine the Mechanism of Action of DNA-Targeted Platinum-Acridine Anticancer Agents

Platinum-based drugs have been used to successfully treat diverse cancers for several decades. Cisplatin, the original compound of this class, cross-links DNA, resulting in cell cycle arrest and cell death <i>via</i> apoptosis. Cisplatin is effective against several tumor types, yet it exhibits toxic side effects and tumors often develop resistance. To mitigate these liabilities while maintaining potency, we generated a library of non-classical platinum-acridine hybrid agents and assessed their mechanisms of action using a validated genome-wide screening approach in <i>Saccharomyces cerevisiae</i> and in the distantly related yeast <i>Schizosaccharomyces pombe.</i> Chemogenomic profiles from both <i>S. cerevisiae</i> and <i>S. pombe</i> demonstrate that several of the platinum-acridines damage DNA differently than cisplatin based on their requirement for distinct modules of DNA repair

Comparative Chemogenomics To Examine the Mechanism of Action of DNA-Targeted Platinum-Acridine Anticancer Agents

Platinum-based drugs have been used to successfully treat diverse cancers for several decades. Cisplatin, the original compound of this class, cross-links DNA, resulting in cell cycle arrest and cell death <i>via</i> apoptosis. Cisplatin is effective against several tumor types, yet it exhibits toxic side effects and tumors often develop resistance. To mitigate these liabilities while maintaining potency, we generated a library of non-classical platinum-acridine hybrid agents and assessed their mechanisms of action using a validated genome-wide screening approach in <i>Saccharomyces cerevisiae</i> and in the distantly related yeast <i>Schizosaccharomyces pombe.</i> Chemogenomic profiles from both <i>S. cerevisiae</i> and <i>S. pombe</i> demonstrate that several of the platinum-acridines damage DNA differently than cisplatin based on their requirement for distinct modules of DNA repair

Comparative Chemogenomics To Examine the Mechanism of Action of DNA-Targeted Platinum-Acridine Anticancer Agents

Platinum-based drugs have been used to successfully treat diverse cancers for several decades. Cisplatin, the original compound of this class, cross-links DNA, resulting in cell cycle arrest and cell death <i>via</i> apoptosis. Cisplatin is effective against several tumor types, yet it exhibits toxic side effects and tumors often develop resistance. To mitigate these liabilities while maintaining potency, we generated a library of non-classical platinum-acridine hybrid agents and assessed their mechanisms of action using a validated genome-wide screening approach in <i>Saccharomyces cerevisiae</i> and in the distantly related yeast <i>Schizosaccharomyces pombe.</i> Chemogenomic profiles from both <i>S. cerevisiae</i> and <i>S. pombe</i> demonstrate that several of the platinum-acridines damage DNA differently than cisplatin based on their requirement for distinct modules of DNA repair

Legislative Documents

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