Figure 5.

Mcm21 is required for kinetochore biorientation when Ipl1 function is impaired. (A) Serial dilutions (fivefold) of WT, mcm21Δ, deg-ipl1, and mcm21Δ deg-ipl1 (SBY818, SBY1897, SBY6940, and SBY5551) cells were plated in the presence or absence of doxycycline. (B) Sister separation of the ChrIV pericentromeric locus was monitored in strains in A released from G1 into a nocodazole arrest in the presence of doxycycline. (C) Lysates from strains in A were immunoblotted against Pds1-myc and tubulin during a synchronous cell cycle. (D) Fivefold dilutions of WT, mcm21Δ, and mcm21Δ mad3Δ cells (SBY818, SBY1897, and SBY7656) were plated for viability. (E) Lysates of cells in D were immunoblotted for Pds1-myc and tubulin during a synchronous cell cycle. (F) ChrIV segregation was assessed in strains in A that reached anaphase during a synchronous cell cycle in the presence of doxycycline (n > 100).