Figure 6
Lack of DNA cleavage activity in C. elegans SPO-11. (a) The indicated concentration of SPO-11 was incubated with negatively supercoiled dsDNA (sc dsDNA) with or without Mg2+, ATP, or Mg2+-ATP as indicated. DNA was released from nucleoprotein complexes by treatment with SDS and proteinase K (PK). (b) Purified SPO-11, human TOP2α, and TOP2β were analyzed by SDS-PAGE. (c) The indicated concentration of SPO-11, TOP2α, or TOP2β was incubated with negatively supercoiled dsDNA in the condition with the presence of Mg2+-ATP. (d) Linear dsDNA was used as the substrate to examine the DNA cleavage activity of SPO-11. (e) SPO-11 was examined for DNA cleavage activity with the indicated metal ion.
