Figure 8.

Analysis of RAM2, a candidate SET1-interacting gene. (A) Transcription of RAM2 and HAS1 was analyzed in SET1 (UCC1001) and set1 (LPY1297) strains by hybridizing blots containing RNA from each strain with probes prepared from radioactively labeled DNA fragments obtained in the target screen. ACT1 served as a loading control. RAM2 and HAS1 transcript levels were diminished in set1 mutant RNA, suggesting that these genes are transcriptional targets of Set1p. (B) Prenyltransferase assay of protein extracts prepared from SET1 (UCC1001) and set1 (LPY1297) mutants. Clarified extracts from duplicate cultures of both strains were analyzed for incorporation of ³H-farnesylpyrophosphate into trichloroacetic acid-precipitable counts as a function of time. Filled squares indicate SET1 extracts, open triangles indicate set1 extracts, and open circles indicate control extracts from SET1 cells plus 5 mM EDTA.