Fig. 4.

Mlo3 associates with the TRAMP complex, which is involved in antisense RNA suppression. (A) Mass spectrometry analysis of Mlo3-Flag or Cid14-Flag purified fractions. A detailed list of the proteins identified is provided in table S2 and S3. (B) TRAMP association with Mlo3 is independent of nucleic acids. IP of Cid14-Flag using antibody to Flag was followed by Western blotting with the indicated antibodies. (C) Hierarchical clustering of mutants on the basis of similarities of their antisense profiles. Pairwise comparisons of anti-sense profiles were performed by calculating the median antisense ratio (mutant/WT) for 842 genes. Pearson’s correlation coefficients were converted into color codes. Except for mlo3-A and clr6-1 mutants, deletion alleles of genes indicated were used. (D) Model showing processing of centromeric and antisense RNA by the coordinatedactionofMlo3,TRAMP, Clr4, RNAi, and the exosome (see also fig. S14). Because low levels of siRNAs are detected in mlo3Δ cells, it is likely that additional mechanisms also target RNAi to centromeric RNAs.