distinct roles of non-canonical poly(a) polymerases in rna metabolism不同的角色非规范保利(a)在rna聚合酶代谢.pdfVIP

Distinct Roles of Non-Canonical Poly(A) Polymerases in RNA Metabolism 1 2 3 3 1 1 Salvatore San Paolo , Stepanka Vanacova , Luca Schenk , Tanja Scherrer , Diana Blank , Walter Keller *, ´ 3* Andre P. Gerber 1 Department of Cell Biology, Biozentrum, University of Basel, Basel, Switzerland, 2 National Center for Biomolecular Research, Masaryk University, Brno, Czech Republic, ¨ ¨ 3 Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland Abstract Trf4p and Trf5p are non-canonical poly(A) polymerases and are part of the heteromeric protein complexes TRAMP4 and TRAMP5 that promote the degradation of aberrant and short-lived RNA substrates by interacting with the nuclear exosome. To assess the level of functional redundancy between the paralogous Trf4 and Trf5 proteins and to investigate the role of the Trf4-dependent polyadenylation in vivo, we used DNA microarrays to compare gene expression of the wild-type yeast strain of S. cerevisiae with either that of trf4D or trf5D mutant strains or the trf4D mutant expressing the polyadenylation- defective Trf4(DADA) protein. We found little overlap between the sets of transcripts with altered expression in the trf4D or the trf5D mutants, suggesting that Trf4p and Trf5p target distinct groups of RNAs for degradation. Surprisingly, most RNAs the expression of which was altered by the trf4 deletion were restored to wild-type levels by overexpression of TRF4(DADA), showing that the polyadenylation activity of Trf4p is dispensable in vivo. Apart from previously reported Trf4p and Trf5p target RNAs, this analysis along with