Yeast Cth2 protein represses the translation of ARE-containing mRNAs in response to iron deficiency
Fig 5
Cth2 represses its own mRNA translation in a Cth2-TZF-dependent manner under iron-limited conditions.
(A) cth1Δcth2Δ mutant cells transformed with plasmids pRS416-Flag2-CTH2 (CTH2) or pRS416-Flag2-CTH2-C190R (CTH2-C190R) were grown at 30°C for 7 hours in SC-Ura with 100 μM BPS (-Fe). Mean values and standard deviation from four independent experiments of steady-state mRNA, proteins and CTH2 translation efficiencies were determined and normalized as in Fig 4A. (B and C) cth1Δcth2Δ mutant cells transformed with pRS416-CTH2 (CTH2) or pRS416-CTH2-C190R (CTH2-C190R) were cultivated as mentioned above. Polysomal fractionation was carried out as described in Materials and Methods. The RNA of individual fractions was extracted and the percentages of CTH2 (B) and ACT1 mRNA (C) were analyzed by RT-qPCR as described in Materials and Methods. Representative data from at least two independent experiments are shown.