Published online 21 June 2004
Nucleic Acids Research, Vol. 32 No. 11 © Oxford University Press 2004; all rights reserved
Efficient downregulation of immunoglobulin µ mRNA with premature translation-termination codons requires the 5'-half of the VDJ exon
Institute of Cell Biology, University of Bern, Baltzerstrasse 4, CH-3012 Bern, Switzerland
* To whom correspondence should be addressed at Institute of Cell Biology, University of Bern, Baltzerstrasse 4, CH-3012 Bern, Switzerland. Tel: +4131 631 4627; Fax: +4131 631 4616; Email: oliver.muehlemann{at}izb.unibe.ch
Received March 4, 2004; Revised and Accepted May 25, 2004
Premature translation-termination codons (PTCs) elicit rapid degradation of the mRNA by a process called nonsense-mediated mRNA decay (NMD). NMD appears to be significantly more efficient for mRNAs of genes belonging to the immunoglobulin superfamily, which frequently acquire PTCs during VDJ rearrangment, than for mRNAs of other genes. To identify determinants for efficient NMD, we developed a minigene system derived from a mouse immunoglobulin µ gene (Ig-µ) and measured the effect of PTCs at different positions on the mRNA level. This revealed that PTCs located downstream of the V–D junction in the VDJ exon of Ig-µ minigenes and of endogenous Ig-µ genes elicit very strong mRNA downregulation, whereas NMD efficiency decreases gradually further upstream in the V segment where a PTC was inserted. Interestingly, two PTCs are in positions where they usually do not trigger NMD (<50 nt from the 3'-most 5' splice site) still resulted in reduced mRNA levels. Using a set of hybrid constructs comprised of Ig-µ and an inefficient substrate for NMD, we identified a 177 nt long element in the V segment that is necessary for efficient downregulation of PTC-containing hybrid transcripts. Moreover, deletion of this NMD-promoting element from the Ig-µ minigene results in loss of strong NMD.
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