Figure 2.

Chromosome Arms as well as Centromeres Generate Physical Cohesion

(A) Asynchronous K18066 cultures having the Z. rouxii recombinase under the control of the Gal1 promoter (Gal1P-Rec) and a 26 kb circular minichromosome containing RS-Cen3-RS (whose centromere can be uncoupled upon recombinase expression) were grown at 30°C in YEPraff; glucose (Cen3 present) or galactose (Cen3 deleted) were added to 2% final concentration for 3 hr. Cells were shifted into YPD for 1.5 hr, followed by nocodazole arrest for 1 hr 40 min.

(B) K18066 cultures having the 26 kb circular minichromosome bearing the RS-Cen3-RS were arrested at 30°C with the α factor pheromone in YEPraff and released for 1 hr in nocodazole. Glucose or galactose were added to 2% final concentration and incubated for an additional 3 hr, maintaining the nocodazole arrest.