8044 Nucleic Acids Research, 2010, Vol. 38, No. 22 Figure 2. qPCR and immunoblotting conﬁrms translational proﬁling. (A) Quantitative reverse-transcription PCR analysis of translation state for qPCR (light grey bars ±SD, n = 3) and array data (black bars) for indicated transcripts (plotted on a log2 scale so that reduced polysome association in mutant cells is shown as a negative number). (B) Top: total protein levels of Sec9p, Taf7p, Taf3p, Cic1p, Lsm8p, Pub1p and Ade2p were analysed by immmunoblot analysis in indicated yeast strains. Experiments were done in triplicate. Loading control Arp2p (Actin Related Protein 2) is shown beneath each. Middle: densitometry analysis for each protein relative to Arp2p is beneath each panel (dark grey bars). Bottom: polysome/monosome [P/M] ratios from translational proﬁling (light grey bars). translational controls for regulating cyclin expression and further suggest that an eIF4E-Caf20p interaction is important for translational repression of CLN3. In metazoa correct intracellular localization of speciﬁc transcripts is important for both early developmental de- cisions and memory functions in neurons. 4E-BPs provide a mechanism to repress translation of delocalized mRNAs e.g. Drosophila 4E-BP Cup acts with Bruno to repress translation of oskar mRNA (30). Polarised growth in budding yeast requires asymmetric localization of components to the growing bud. Several mRNAs have also been found to be bud-localized. A survey of yeast genes uncovered 22-bud localized transcripts, of which 10 were asymmetrically localized to the bud (31). Intriguingly, six of these 10 genes (EGT2, IST2, MMR1, SRL1, TCB3 and TPO1) are translationally up-regulated in caf20D cells. GO Slim mapping analysis suggests that bud, cell wall, and site of polarized cell growth-localized proteins are among those enriched in the caf20D data set. Together, these observations suggest that Caf20p may