NMD is essential for hematopoietic stem and progenitor cells and for eliminating by-products of programmed DNA rearrangements
- Joachim Weischenfeldt1,2,3,
- Inge Damgaard1,2,3,
- David Bryder4,
- Kim Theilgaard-Mönch1,2,3,
- Lina A. Thoren5,
- Finn Cilius Nielsen2,
- Sten Eirik W. Jacobsen5,
- Claus Nerlov5,6, and
- Bo Torben Porse1,2,3,7
- 1 Section for Gene Therapy Research, Copenhagen University Hospital, 2100 Copenhagen, Denmark;
- 2 Department of Clinical Biochemistry, Copenhagen University Hospital, 2100 Copenhagen, Denmark;
- 3 Biotech Research and Innovation Centre (BRIC), University of Copenhagen, 2200 Copenhagen, Denmark;
- 4 Institute for Experimental Medical Science, BMCI13, Lund University, 22184 Lund, Sweden;
- 5 Hematopoietic Stem Cell Laboratory, Lund Strategic Research Centre for Stem Cell Biology and Therapy, Lund University, 22184 Lund, Sweden;
- 6 European Molecular Biology Laboratory (EMBL) Mouse Biology Unit, 00015 Monterotondo, Italy
Abstract
Nonsense-mediated mRNA decay (NMD) is a post-transcriptional surveillance process that eliminates mRNAs containing premature termination codons (PTCs). NMD has been hypothesized to impact on several aspects of cellular function; however, its importance in the context of a mammalian organism has not been addressed in detail. Here we use mouse genetics to demonstrate that hematopoietic-specific deletion of Upf2, a core NMD factor, led to the rapid, complete, and lasting cell-autonomous extinction of all hematopoietic stem and progenitor populations. In contrast, more differentiated cells were only mildly affected in Upf2-null mice, suggesting that NMD is mainly essential for proliferating cells. Furthermore, we show that UPF2 loss resulted in the accumulation of nonproductive rearrangement by-products from the Tcrb locus and that this, as opposed to the general loss of NMD, was particularly detrimental to developing T-cells. At the molecular level, gene expression analysis showed that Upf2 deletion led to a profound skewing toward up-regulated mRNAs, highly enriched in transcripts derived from processed pseudogenes, and that NMD impacts on regulated alternative splicing events. Collectively, our data demonstrate a unique requirement of NMD for organismal survival.
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Footnotes
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↵7 Corresponding author.
↵7 E-MAIL porse{at}rh.dk; FAX 45-3532-5669.
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Supplemental material is available at http://www.genesdev.org.
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Article is online at http://www.genesdev.org/cgi/doi/10.1101/gad.468808.
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- Received December 21, 2007.
- Accepted March 20, 2008.
- Copyright © 2008, Cold Spring Harbor Laboratory Press