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Nucleic Acids Research Advance Access originally published online on April 27, 2008
Nucleic Acids Research 2008 36(10):3374-3388; doi:10.1093/nar/gkn108
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Nucleic Acids Research, 2008, Vol. 36, No. 10 3374-3388
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA

A role for Caf1 in mRNA deadenylation and decay in trypanosomes and human cells

Angela Schwede1, Louise Ellis2, Julia Luther3, Mark Carrington2, Georg Stoecklin3,4 and Christine Clayton1,4,*

1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany, 2Department of Biochemistry, 80 Tennis Court Rd., Cambridge CB2 1GA, UK, 3German Cancer Research Centre (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany and 4DKFZ-ZMBH Allianz

*To whom correspondence should be addressed. Tel: +49 6221 546 876; Fax: +49 6221 545 894; Email: cclayton{at}zmbh.uni-heidelberg.de

Received October 15, 2007. Revised February 25, 2008. Accepted February 26, 2008.

The eukaryotic Ccr4/Caf1/Not complex is involved in deadenylation of mRNAs. The Caf1 and Ccr4 subunits both potentially have deadenylating enzyme activity. We investigate here the roles of Ccr4 and Caf1 in deadenylation in two organisms that separated early in eukaryotic evolution: humans and trypanosomes. In Trypanosoma brucei, we found a complex containing CAF1, NOT1, NOT2 and NOT5, DHH1 and a possible homologue of Caf130; no homologue of Ccr4 was found. Trypanosome CAF1 has deadenylation activity, and is essential for cell survival. Depletion of trypanosome CAF1 delayed deadenylation and degradation of constitutively expressed mRNAs. Human cells have two isozymes of Caf1: simultaneous depletion of both inhibited degradation of an unstable reporter mRNA. In both species, depletion of Caf1 homologues inhibited deadenylation of bulk RNA and resulted in an increase in average poly(A) tail length.


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