Fig 8. Rif2 is recruited to DNA ends, and its lack enhances MRX and MR^V1269MX association to the DSB.

(A, B) ChIP analysis. Exponentially growing YEPR cell cultures were transferred to YEPRG at time zero. Relative fold enrichment of Mre11-Myc fusion protein at the indicated distances from the HO cleavage site was determined after ChIP with anti-Myc antibodies and subsequent qPCR analysis. Plotted values are the mean values with error bars denoting s.d. (n = 3). (C) Flag-tagged Tel1 (50 ng) was incubated with MR^V1269MX (100 ng) in the absence or presence of Rif2 (100 ng), and protein complexes were captured by anti-Flag resin and followed by immunoblotting analysis. (D) Rif2 (150, 300 and 600 nM) was incubated with ³²P-labeled 100-bp dsDNA (10 nM) in the presence of ATP. In lane 5, the reaction mixture was deproteinized with SDS and proteinase K (PK) prior to analysis. Plotted values are the mean value with error bars denoting s.d. (n = 3). (E) ChIP analysis. As in (A), but showing Rif2 recruitment at the HO-induced DSB.