Skip to main content
. 2012 Jun 12;10(6):e1001342. doi: 10.1371/journal.pbio.1001342

Figure 1. Tethered Dhh1 represses translation independent of decapping.

Figure 1

(A) Diagram of reporter mRNAs Dhh1 was tethered to. Each reporter was expressed under control of the GAL1 UAS, and each reporter has two MS2 binding stem-loops engineered in its 3′ UTR. First reporter, MFA2; second reporter, M/GFP; third reporter, P/GFP. Transcriptional shut-off analysis of MFA2 in either wild-type cells (B) or dcp2Δ cells (C) expressing either MS2 alone or tethered Dhh1. RNA was isolated from cells collected at each time point and Northern blot for the reporter was performed. Blots were stripped and reprobed for SCR1 as a loading control. Half-lives are reported in minutes to the right of the gels. (D) Western blot analysis of GFP from either M/GFP or P/GFP co-expressed with either MS2 alone or tethered Dhh1 in wild-type cells or dcp2Δ cells. Blots were stripped and reprobed for Pgk1 as a loading control. (E) Relative quantitation of GFP protein signal normalized to Pgk1 protein signal from Figure 1D. For a given experiment, signal with MS2 alone tethered was set to 100% and signal with Dhh1 tethered was expressed as a percentage of tethering MS2 alone.