FIG. 7.

Hhp1p is hyperphosphorylated in starved cells. (A) The amount of Hhp1p remains the same upon starvation. Total acid-soluble macronuclear protein, extracted from vegetatively growing (GR) or 24-h-starved (ST) wild-type cells, was resolved on an SDS–12% polyacrylamide gel, blotted onto nitrocellulose, and immunoblotted with α-Hhp1p antibodies. (B) Proteins from growing or starved wild-type cells were resolved on an acid-urea (AU) gel and analyzed by Western blot analysis with α-Hhp1p antibodies. In the sample shown in lane 3, Hhp1p from starved cells was incubated with alkaline phosphatase (+AP) before electrophoresis on a long acid-urea gel. Note the appearance of two slower-migrating bands in the sample from starved cells (indicated by arrowheads in lane 2).