FIG. 9.

Cdc13ΔDBp overproduction delays the relocalization of SIN components. (A and B) Cdc7p remains on both SPBs under conditions of high Cdc2p kinase activity. cdc7-HA cells with an integrated copy of cdc13ΔDB under the control of the nmt1 promoter (KGY2474) were grown at 32°C in the presence (A) or absence (B) of thiamine, and samples were collected 18 h later. Cells were fixed in methanol and stained with anti-HA (12CA5) monoclonal antibodies followed by Alexa-conjugated goat anti-mouse secondary antibodies. Rows: a, cells in metaphase or early anaphase; b to d, cells in later stages of anaphase. (C and D) Cells producing endogenously tagged GFP-Sid1p and containing the integrated copy of cdc13ΔDB were grown in the presence (C) or absence (D) of thiamine for 18 h and fixed in methanol, and the localization of GFP-Sid1p and DNA was examined. (E) Cells producing endogenously tagged Sid2p-GFP and containing an integrated copy of cdc13ΔDB were grown in the presence or absence of thiamine for 18 h, and the localization of Sid2p-GFP was examined. Rows: a and b, cells in metaphase; c and d, cells in anaphase.