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Figure 1 | Journal of Molecular Signaling

Figure 1

From: PNRC is a unique nuclear receptor coactivator that stimulates RNA polymerase III-dependent transcription

Figure 1

PNRC interacts with a subunit of RNA pol III. A, ADGal4-RPC39212–316 expression plasmid was isolated from a human mammary gland expression library screening using DBDGal4-PNRC270–327 as bait. To confirm the interaction and the specificity of the interaction between PNRC and RPC39, yeast strain Y187 was cotransformed with the expression plasmids for the expression of the fusion proteins as indicated, and transformants containing these plasmids were selected by growth on SD/-Leu/-Trp agar plates. The expression of interacting hybrid proteins in Y187 transformants was analyzed for LacZ expression. DBDGal4 (DBD) and DBDGal4-human lamin C (hLC) expression plasmids were included as background and negative control, respectively. β-Galactosidase activities in liquid cultures are expressed in Miller units as mean ± s.d. of three independent assays. B, PNRC interacts with the C-terminus, amino acids 212–316, of the human Pol III subunit RPC39. The yeast expression plasmids, pACT2-RPC39 and pACT2-RPC391–212, for the expression of ADGal4-RPC39 wild type (WT) or ADGal4-RPC1–212 (1–212) fusion proteins were constructed as described in 'Methods'. The expression plasmids for ADGal4-RPC39212–316 (212–316) and ADGal4-RPC39241–316(241–316) were isolated from library screening through their interaction with PNRC270–327. Y187 cells were cotransformed with the expression plasmids coding for the fusion proteins as indicated. The selection of Y187 transformants and the β-Gal assays on transformants were performed as described in A.

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