FIGURE 6.

The roles of CCR4-NOT complex in TNRC6A-mediated translation repression in yeast. A–C, translation repression by tethered TNRC6A was almost diminished in ccr4Δ and caf1Δ mutant cells independent of a poly(A) tail. Cell extracts were prepared from indicated cells harboring the indicated plasmid, and polysome analysis was performed. RNA samples were analyzed by Northern blotting using a DIG-labeled PGK1 3′-UTR probe. D, the expression level of Not1 reduced in caf1Δ and ccr4Δ mutant cells. Endogenous NOT1, CAF1, and CCR4 were tagged by 3× hemagglutinin (HA) in wild type, caf1Δ, and ccr4Δ mutant cells. Protein samples were analyzed by Western blotting using anti-HA, anti-Pab1 and anti-eEF1α antibodies.