Table II

cdc12p Localization Patterns in Wild-type and Mutant Cells

Table II
Percentage of cells with particular cdc12p localization pattern
Cell typeNumber of cells assayed
%
Wild type  interphase300100 0 0 0 0 0 0 0
Wild type  early mitosis 50 52 0 048 0 0 0 0
Wild type  late mitosis 80  2 0 098 0 0 0 0
OP cdc12  interphase205 3337 0 01812 0 0
OP cdc12  early mitosis 70  1173150 0 0 0 0
OP cdc12  late mitosis 80  1 0 099 0 0 0 0
cdc8 mitosis 50100 0 0 0 0 0 0 0
cdc15 mitosis 50 88 0 0 6 0 0 0 0
cdc4 early mitosis 46 7613 011 0 0 0 0
cdc4 late mitosis 46 1576 6 2 0 0 0 0
mid1 early mitosis 70 3916i 0 0 0 0 343
mid1 late mitosis 70  4 3 013 0 03446
  • Cells were fixed and stained with anti-cdc12 and anti-tubulin antibodies and DAPI. Individual cells were first screened by fluorescence microscopy for a particular cell cycle stage on the basis of tubulin staining, and were then examined for cdc12p localization. Cells with a short mitotic spindle before nuclear division were designated as being in early mitosis. Cells in anaphase or postanaphase were designated as being in late mitosis. Cells with an interphase microtubule array and a single nucleus were designated as being in interphase. Dots and lines indicate cdc12p localization, and the empty circle indicates the nucleus. OP (overproducing) cdc12: cells (FC566) that contain a chromosomal integrated copy of pnmt-cdc12 were induced in minimal media for 20 h at 30°C. cdc8-346 (FC131), cdc15-287 (FC55), and cdc4-377 (FC361) mutants were grown for 4 h at the restrictive temperature of 35.5°C. mid1-366 (FC164) cells were grown for 2 h at 35.5°C. (a) Cells with no specific cdc12p staining. (b) Cells with a medial cdc12p spot. Interphase spots were adjacent to the nucleus. (c) Cells with a cdc12p spot with an intersecting ring. (d) Cells with a normal cdc12p ring. (e) Cells with a cdc12p dot near the end of the cell, which probably is a midbody remnant, left from a previous division. (f) Cells with multiple dots in overexpressing cells or dots not covered in b or e. Many cells in this category have a combination of patterns seen in b and e. (g) Cells with an intact but displaced ring. (h) Cells with a cdc12p strand. Many are associated with a dot, and >90% are associated with the nucleus. (i) In mid1 cells, dots are not always on the cortex, and some dots (∼30%) are displaced away from the medial nucleus.