Category:Team On Holliday

Cdk1 activates Separase (Esp1 in yeast) through phosphorylation to trigger anaphase onse. In figure 1D, Esp1 phosphorylation was detected after around 30 minutes, before the major mitotic cyclin-Cdk complex in yeast (CDK1-Clb2) started to appear in vivo.

Figure 2 provides the biochemical properties of the RFS-1/RIP-1 complex. The western blot in part A of this figure confirms the precence of the two proteins (RFS-1/RIP-1) after co-purifying the recombinant proteins from budding yeast cells. The Electrophoretic mobility shift assays (EMSA) shown in part B of the figure reveals that the RFS-1/RIP-1 complex binds in a nucleotide independent manner weakly to ssDNA but not dsDNA. Bands are observed only for ssDNA. The RFS-1/RIP-1 complex can also stimulate D loop formations with RAD51 (C) and the fact that ATP is required to do so (D).

Figure 1 depicts the cessation of the dissociation of RAD-51 from ssDNA due to the presence of protein RFS-1/RIP-1. Figure 1, Part H [(i), (ii)] shows the known mechanism of RAD-51 by demonstrating that it’s presence prohibits the binding of ScRPA-eGFP-ssDNA. The incapability of RAD-51 to be expressed due to the lack of ATP is found in Figure 1, Part H (ii). Figure 1 ultimately illustrates that in the precence of the protein RFS-1/RIP-1 and ATP; RAD-51 along with the protein RFS-1/RIP-1 is present but the ScRPA-eGFP-ssDNA are not [(Figure 1, Part H (iii)]. The same events occur when the protein RFS-1/RIP-1 is present but not ATP [(Figure 1, Part H (iv)].