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. 2000 Nov 28;97(25):13732–13737. doi: 10.1073/pnas.250400997

Figure 4.

Figure 4

The interaction between Nhp6B and Tup1p is biologically relevant. (A) Nhp6 is necessary for glucose repression of the GAL1 promoter. RNA was prepared from the depicted strains carrying a GAL1-LacZ fusion integrated at the GAL1 locus. JD53 was used as wild-type parental strain (lanes 1 and 4). The ΔNHP6 strain was derived from JD53 that lacks NHP6A and NHP6B (lanes 2 and 5). In the strain ΔNHP6 + NHP6 (lanes 3 and 6), NHP6A and NHP6B had been reintegrated into the original loci. Equal amounts of total RNA were loaded as confirmed by ethidium bromide staining (not shown) and background hybridization to the 28 S rRNA (Right). The Northern blot was probed with a LacZ probe (lanes 1–3) and with an ACT1 probe (lanes 4–6). We consistently saw a slight increase in the level of ACT1 mRNA in the ΔNHP6 strain. (B) Nhp6 is not necessary for α2p repression. RNA was prepared from the depicted strains, and the Northern blot was probed with an MFA1 probe (Upper) or with an ACT1 probe (Lower). In lane 1, RNA was isolated from JD52, a MATa strain. In lane 2, RNA was isolated from JD53, which was used as wild-type parental MATα strain. Lane 3 contained RNA from JD53 lacking NHP6A and NHP6B (ΔNHP6). For lane 4, NHP6A and NHP6B had been reintegrated into the original loci (ΔNHP6 + NHP6). Lane 5 contained RNA from JD53 lacking TUP1 (ΔTUP1). (C) NHP6 and REG1 deletions are synthetically lethal. Shown are serial dilutions of the depicted S. cerevisiae strains carrying a URA3-marked Nhp6B expression plasmid (YCplac33-NHP6B) on medium lacking or containing FOA.