Journal: Cancer Research

Article Title: Chromosome Breakage Is Regulated by the Interaction of the BLM Helicase and Topoisomerase IIα

doi: 10.1158/0008-5472.can-10-1727

Figure Lengend Snippet: Figure 2. BLM helicase activity is enhanced by topoisomerase IIa using 3’ overhang and bubble, but not X-junction, substrates. A, Coomassie blue–stained polyacrylamide gel of purified recombinant 6xHis-tagged BLM (100 ng) used in helicase assays. B, graphical representation of percent unwinding of the 30

Article Snippet: Western blotting used antibodies for BLM, topoisomerase IIa (Ab-1; Calbiochem), or topoisomerase IIIa (N-20; Santa Cruz Biotechnology).

Techniques: Activity Assay, Staining, Purification, Recombinant

Journal: Cancer Research

Article Title: Chromosome Breakage Is Regulated by the Interaction of the BLM Helicase and Topoisomerase IIα

doi: 10.1158/0008-5472.can-10-1727

Figure Lengend Snippet: Figure 5. Amino acids 489–587 in BLM are necessary for enhancing BLM unwinding by topoisomerase IIa. A, autoradiograph of time-course helicase assays with 2 fmol of 30 overhang substrate and indicated amounts of BLM or BLMD489–587 (left) or wild-type BLM (right) with and without topoisomerase IIa. Quantification of percent unwinding by BLMD489–587 or wild-type BLM with equimolar topoisomerase IIa is shown. Only wild-type BLM is stimulated by topoisomerase IIa. Minus ATP (ATP) control indicates ATP-dependent unwinding of substrate and (*) depicts completely unwound heat-denatured substrate. B, increasing concentrations of topoisomerase IIa (0–10 nmol/L) do not stimulate BLMD489–587 but stimulate wild-type BLM. Helicase assays were carried out with different concentrations of topoisomerase IIa and a fixed concentration of BLM (1 nmol/L) for 10 minutes. TOPO, topoisomerase.

Article Snippet: Western blotting used antibodies for BLM, topoisomerase IIa (Ab-1; Calbiochem), or topoisomerase IIIa (N-20; Santa Cruz Biotechnology).

Techniques: Autoradiography, Control, Concentration Assay