Cytoplasmic foci are sites of mRNA decay in human cells

doi:10.1083/jcb.200309008

Published online 5 April 2004. doi:10.1083/jcb.200309008
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 165, Number 1, 31-40

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Article

Cytoplasmic foci are sites of mRNA decay in human cells

Nicolas Cougot, Sylvie Babajko, and Bertrand Séraphin

Équipe labellisée La Ligue, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France

Address correspondence to B. Séraphin, Équipe labellisée La Ligue, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, Avenue de la Terrasse, 91198 Gif sur Yvette, France. Tel.: 33 1 69 82 38 84. Fax: 33 1 69 82 38 77. email: seraphin{at}cgm.cnrs-gif.fr

Understanding gene expression control requires defining themolecular and cellular basis of mRNA turnover. We have previouslyshown that the human decapping factors hDcp2 and hDcp1a areconcentrated in specific cytoplasmic structures. Here, we showthat hCcr4, hDcp1b, hLsm, and rck/p54 proteins related to 5'–3'mRNA decay also localize to these structures, whereas DcpS,which is involved in cap nucleotide catabolism, is nuclear.Functional analysis using fluorescence resonance energy transferrevealed that hDcp1a and hDcp2 interact in vivo in these structuresthat were shown to differ from the previously described stressgranules. Our data indicate that these new structures are dynamic,as they disappear when mRNA breakdown is abolished by treatmentwith inhibitors. Accumulation of poly(A)⁺ RNA in these structures,after RNAi-mediated inactivation of the Xrn1 exonuclease, demonstratesthat they represent active mRNA decay sites. The occurrenceof 5'–3' mRNA decay in specific subcellular locationsin human cells suggests that the cytoplasm of eukaryotic cellsmay be more organized than previously anticipated.

Key Words: cell cycle; FRET; green fluorescent protein; transcriptional and translational inhibitors; RNA stability

Abbreviations used in this paper: FRET, fluorescence resonanceenergy transfer; NMD, nonsense-mediated decay; NSD, nonstopdecay.

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