Nucleic Acids Research Advance Access originally published online on July 13, 2009
Nucleic Acids Research 2009 37(16):5511-5528; doi:10.1093/nar/gkp571
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Nucleic Acids Research, 2009, Vol. 37, No. 16 5511-5528
© 2009 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.
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The role of deadenylation in the degradation of unstable mRNAs in trypanosomes
Zentrum für Molekulare Biologie (ZMBH), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
*To whom correspondence should be addressed. Tel: 49 6221 546 876; Fax: 49 6221 545 894; Email: cclayton{at}zmbh.uni-heidelberg.de
Received May 15, 2009. Revised June 16, 2009. Accepted June 20, 2009.
Removal of the poly(A) tail is the first step in the degradation of many eukaryotic mRNAs. In metazoans and yeast, the Ccr4/Caf1/Not complex has the predominant deadenylase activity, while the Pan2/Pan3 complex may trim poly(A) tails to the correct size, or initiate deadenylation. In trypanosomes, turnover of several constitutively-expressed or long-lived mRNAs is not affected by depletion of the 5'–3' exoribonuclease XRNA, but is almost completely inhibited by depletion of the deadenylase CAF1. In contrast, two highly unstable mRNAs, encoding EP procyclin and a phosphoglycerate kinase, PGKB, accumulate when XRNA levels are reduced. We here show that degradation of EP mRNA was partially inhibited after CAF1 depletion. RNAi-targeting trypanosome PAN2 had a mild effect on global deadenylation, and on degradation of a few mRNAs including EP. By amplifying and sequencing degradation intermediates, we demonstrated that a reduction in XRNA had no effect on degradation of a stable mRNA encoding a ribosomal protein, but caused accumulation of EP mRNA fragments that had lost substantial portions of the 5' and 3' ends. The results support a model in which trypanosome mRNAs can be degraded by at least two different, partially independent, cytoplasmic degradation pathways attacking both ends of the mRNA.
Present addresses: Natalia Bercovich, Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI), FCEyN, Dto. FBMC, Universidad de Buenos Aires, Vuelta de Obligado 2490 2P, 1428, Buenos Aires, Argentina.
Mhairi Stewart, Glasgow Biomedical Research Centre, University of Glasgow, G12 8QQ, UK.