FIG. 1.

A 9MYC-DIA2 strain behaves like the wild type. (A) Growth of the Dia2^Myc strain is indistinguishable from that of the wild type on rich medium with the indicated amounts of HU or methyl methanesulfonate (MMS) at room temperature. Tenfold serial dilutions are shown. (B) Dia2^Myc associates with Skp1. Skp1 was immunoprecipitated with anti-Skp1 antibodies (36) from an untagged strain or 9MYC-DIA2 strains (lanes 3 and 4, respectively). Immunoblots were probed with anti-myc or anti-Skp1 antibodies. The bottom panel shows that Dia2^Myc is immunoprecipitated (IP) with anti-Skp1 antibodies but not with preimmune serum (PI) or protein A/G beads (Beads) in the absence of antibody. (C) A fraction of Dia2^Myc is chromatin bound. Dia2^Myc cells were fractionated as described previously (30a). Crude-pellet-associating Dia2^Myc is released with DNase I (compare lanes 3 and 5) or high-salt (compare lanes 3 and 7) treatment. Mcm4^HA was monitored as a positive control. WCE, whole-cell extract; Su, supernatant; Pel, pellet. Total protein was stained using Ponceau S to visualize overall protein fractionation from equal loaded volumes.