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| Subject Categories: RNA |
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| The EMBO Journal
(2002) 21, 1427–1436, doi: 10.1093/emboj/21.6.1427 |
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| Ccr4p is the catalytic subunit of a Ccr4p/Pop2p/Notp mRNA deadenylase complex in Saccharomyces cerevisiae |
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| Morgan Tucker, Robin R. Staples, Marco A. Valencia-Sanchez, Denise Muhlrad and Roy Parker |
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Department of Molecular and Cellular Biology and Howard Hughes Medical Institute, 1007 East Lowell Street, University of Arizona, Tucson, AZ 85721, USA To whom correspondence should be addressed Roy Parker, rrparker@u.arizona.edu Received 17 October 2001; Revised 10 December 2001; Accepted 19 December 2001. |
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| Abstract |
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| The major pathways of mRNA turnover in eukaryotic cells are initiated by shortening of the poly(A) tail. Recent work has identified Ccr4p and Pop2p as components of the major cytoplasmic deadenylase in yeast. We now demonstrate that CCR4 encodes the catalytic subunit of the deadenylase and that Pop2p is dispensable for catalysis. In addition, we demonstrate that at least some of the Ccr4p/Pop2p-associated Not proteins are cytoplasmic, and lesions in some of the NOT genes can lead to defects in mRNA deadenylation rates. The Ccr4p deadenylase is inhibited in vitro by addition of the poly(A) binding protein (Pab1p), suggesting that dissociation of Pab1p from the poly(A) tail may be rate limiting for deadenylation in vivo. In addition, the rapid deadenylation of the COX17 mRNA, which is controlled by a member of the Pumilio family of deadenylation activators Puf3p, requires an active Ccr4p/Pop2p/Not deadenylase. These results define the Ccr4p/Pop2p/Not complex as the cytoplasmic deadenylase in yeast and identify positive and negative regulators of this enzyme complex. |
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| Keywords: deadenylation, decay, mRNA, turnover, yeast |
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