FIGURE 5.

Endogenous sequences induce translation arrest coupled with protein degradation. A, construction of the pGPDp-GFP-X-FLAG-HIS3 reporter plasmids. These plasmids contain various sequences inserted between GFP and HIS3. The inserted amino acid sequences and original gene names are shown. B, endogenous sequence elements induce translation arrest coupled with protein degradation by the proteasome. W303 cells were transformed with pGPDp-GFP-X-FLAG-HIS3 plasmids that contained sequences from the indicated genes. The cells were grown in SC-Ura, and the samples were analyzed using Western blotting with anti-GFP antibodies. When indicated (+), cell extracts were prepared 2 h after the addition of 0.2 mm MG132. C, translation arrest products in the not4Δ mutant. W303not4Δ cells were transformed with pGPDp-GFP-X-FLAG-HIS3 plasmids that contained sequences from the indicated genes. The cells were grown in SC-Ura, and the samples were analyzed using Western blotting with anti-GFP antibodies.