Figure 6.

The translation of the N-terminal region of Asn1 is required to recruit Tma108 on the ribosome. Real-time quantitative RT-PCR analyses of the enrichment in Tma108-PA IPs of various ASN1 versions fused to a 13-myc Tag (A, construct 1–3) or a LacZ gene (B, construct 4–5). For construct 5, the experiment was also performed with a mutated version of the Tma108-PA (E296Q) in which the glutamate of the MAMEN motif was replaced by a glutamine. The arrows in bold indicate the positions of the primers used for the Q-PCR. The mean enrichment factors were obtained from two independent immunoprecipitation experiments by calculating the ratio IP/Input normalized using ACT1 and ATP1 as control mRNAs. In each experiment, the enrichment of the endogenous ATP2 mRNA was measured and confirmed that Tma108-PA ribonucleoparticles were efficiently immunoprecipitated (data not shown).