RNAi-dependent and -independent RNA turnover mechanisms contribute to heterochromatic gene silencing.
In fission yeast, the RNAi pathway is required for heterochromatin-dependent silencing of transgene insertions at centromeric repeats and acts together with other pathways to silence transgenes at the silent mating-type locus. Here, we show that transgene transcripts at centromeric repeats are processed into siRNAs and are therefore direct targets of RNAi. Furthermore, we show that Cid14, a member of the Trf4/5 family of poly(A) polymerases, has poly(A) polymerase activity that is required for heterochromatic gene silencing. Surprisingly, while siRNA levels in cid14Delta cells are dramatically reduced, the structural integrity of heterochromatin appears to be preserved. Cid14 resides in a complex similar to the TRAMP complex found in budding yeast, which is part of a nuclear surveillance mechanism that degrades aberrant transcripts. Our findings indicate that polyadenylation by a TRAMP-like complex contributes to robust silencing of heterochromatic genes in fission yeast via the recruitment of the exosome and/or the RNAi machinery.
Bühler
Marc
M
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Haas
Wilhelm
W
Gygi
Steven P
SP
Moazed
Danesh
D
D015536:Down-Regulation / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D020868:Gene Silencing / Q000502:physiology*; D006570:Heterochromatin / Q000235:genetics*; D046911:Macromolecular Substances / Q000378:metabolism; D011062:Polynucleotide Adenylyltransferase / Q000235:genetics*; D034622:RNA Interference / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics*
D006570:Heterochromatin; D046911:Macromolecular Substances; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D029702:Schizosaccharomyces pombe Proteins; C509157:Cid14 protein, S pombe; D011062:Polynucleotide Adenylyltransferase