TY - DATA T1 - Cohesin rings devoid of Scc3 and Pds5 topologically embrace circular DNA. PY - 2012/08/09 AU - Irina Kulemzina AU - Martin R. Schumacher AU - Vikash Verma AU - Jochen Reiter AU - Janina Metzler AU - Antonio Virgilio Failla AU - Christa Lanz AU - Vipin T. Sreedharan AU - Gunnar Rätsch AU - Dmitri Ivanov UR - https://plos.figshare.com/articles/figure/_Cohesin_rings_devoid_of_Scc3_and_Pds5_topologically_embrace_circular_DNA_/264717 DO - 10.1371/journal.pgen.1002856.g008 L4 - https://ndownloader.figshare.com/files/594235 KW - rings KW - devoid KW - scc3 KW - pds5 KW - topologically KW - circular N2 - Strains 1021 (untagged), 1813 (SCC3-HA6, SCC1-Myc18), 1625 (SCC3-HA6-degron, SCC1-Myc18), 2525 (PDS5-HA6, SCC1-Myc18) and 1818 (PDS5-HA6-degron, SCC1-Myc18) carried the centromeric minichromosomes. (A) Yeast lysates were incubated with BglII restriction enzyme as indicated. Minichromosomes were co-immunoprecipitated with Scc1-Myc18. DNA was prepared by phenol/chloroform extraction and separated on a 1% agarose gel with ethidium bromide. Southern blot probed with a TRP1-specific probe is shown. Nicked (N), linear (L), and closed circular (C) forms of the minichromosome are indicated. (B) Minichromosomes were immunoprecipitated with anti-HA antibody. Minichromosomes from SCC3-HA6 but not SCC3-HA6-degron strains could be co-immunoprecipitated with Scc3 indicating the efficient depletion of Scc3 from the minichromosomes in the SCC3-HA6-degron strain. Since Pds5 association with minichromosomes is very salt-sensitive, they could not be co-immunoprecipitated with Pds5-HA6 in either the wild type or PDS5-HA6-degron strains under our experimental conditions. ER -