Understanding gene expression control requires defining the molecular and cellular basis of mRNA turnover. We have previously shown that the human decapping factors hDcp2 and hDcp1a are concentrated in specific cytoplasmic structures. Here, we show that 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 transfer revealed that hDcp1a and hDcp2 interact in vivo in these structures that were shown to differ from the previously described stress granules. Our data indicate that these new structures are dynamic, as they disappear when mRNA breakdown is abolished by treatment with inhibitors. Accumulation of poly(A)+ RNA in these structures, after RNAi-mediated inactivation of the Xrn1 exonuclease, demonstrates that they represent active mRNA decay sites. The occurrence of 5′–3′ mRNA decay in specific subcellular locations in human cells suggests that the cytoplasm of eukaryotic cells may be more organized than previously anticipated.
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12 April 2004
Article|
April 05 2004
Cytoplasmic foci are sites of mRNA decay in human cells
In Special Collection:
JCB65: RNA
Nicolas Cougot,
Nicolas Cougot
Équipe labellisée La Ligue, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France
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Sylvie Babajko,
Sylvie Babajko
Équipe labellisée La Ligue, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France
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Bertrand Séraphin
Bertrand Séraphin
Équipe labellisée La Ligue, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France
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Nicolas Cougot
Équipe labellisée La Ligue, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France
Sylvie Babajko
Équipe labellisée La Ligue, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France
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: [email protected]
Abbreviations used in this paper: FRET, fluorescence resonance energy transfer; NMD, nonsense-mediated decay; NSD, nonstop decay.
Received:
September 02 2003
Accepted:
March 05 2004
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2004
J Cell Biol (2004) 165 (1): 31–40.
Article history
Received:
September 02 2003
Accepted:
March 05 2004
Citation
Nicolas Cougot, Sylvie Babajko, Bertrand Séraphin; Cytoplasmic foci are sites of mRNA decay in human cells . J Cell Biol 12 April 2004; 165 (1): 31–40. doi: https://doi.org/10.1083/jcb.200309008
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