Figure 3.

The cohesin loader Mis4/Ssl3 partly colocalizes with cohesin at tRNA and ribosomal protein genes. (a) Comparison of Rad21, Mis4, Ssl3, Cnd2, Sfc6 and Fhl1 localization, all fused to a Pk₉ epitope tag, by chromatin immunoprecipitation. Cnd2 was analyzed in an nda3-KM311 strain arrested in mid-mitosis at the restrictive temperature, and the other proteins were analyzed in exponentially proliferating cells. A 100 kb region of chromosome 2 is shown. Grey bars above and below the midline represent ORFs transcribed from left to right and opposite, respectively. tRNA and ribosomal protein genes are highlighted in red. (b) Cytological colocalization analysis. Spread chromosomes were stained with antibodies against the Pk epitope tag, and against GFP, to detect the indicated pairs of proteins. DNA was counterstained with DAPI.