FIG. 1.
Effects of deletion of the PCL8 and/or PCL10 gene on yeast glycogen metabolism. Wild-type (wt), pho85, pcl8, pcl10, and pcl8 pcl10 yeast strains, as indicated, were analyzed as follows. (A) Glycogen levels were measured as described in Materials and Methods. The strains used were EG328-1A (wt), DH28 (pho85), DH96-11 and DH96-52 (pcl8), DH93-81 and DH93-82 (pcl10), and DH97-13 and DH97-33 (pcl8 pcl10). Averages and standard errors of three independent experiments are shown. (B) Glycogen synthase −/+ glucose-6-P activity measured in extracts from the strains represented in panel A. Averages and standard errors of three independent experiments are shown. (C) Glycogen synthase kinase activity was measured by the ability of cell extracts to transfer 32Pi from ATP to added purified Gsy2, which was then subjected to SDS-PAGE. Shown is an autoradiogram from one of three experiments yielding similar results. The relevant genotypes are indicated, and the strains analyzed were EG328-1A (wt), DH96-52 (pcl8), DH93-82 (pcl10), DH97-13 (pcl8 pcl10), and DH35-64 (pho85). Gsy2 refers to a control lacking added yeast extract. The molecular masses of standards are indicated, in kilodaltons.