Alternative splicing induced by nonsense mutations in the immunoglobulin μ VDJ exon is independent of truncation of the open reading frame

Abstract

In addition to triggering nonsense-mediated mRNA decay (NMD), premature translation-termination codons (PTCs) frequently induce alternative splicing, an observation referred to as nonsense-associated alternative splicing (NAS). In many cases, NAS is induced because the nonsense mutation alters a splicing signal, such as inactivating an exonic splicing enhancer. However, for a few genes, NAS was reported to be PTC specific, implying that a translation signal could influence splicing. Here, we investigated whether production of a previously undetected alternatively spliced transcript from immunoglobulin μ (Ig-μ) depends on premature termination of the open reading frame. We show that PTCs at different positions in the VDJ exon of an Ig-μ minigene activate usage of an alternative 3′ splice site, generating an alternative transcript that lacks the initial PTC and a previously identified NMD-promoting element (NPE), but contains new PTCs because of a frame shift. Corroborating the importance of the NPE for maximal NMD response, the alternative transcript is only moderately down-regulated by NMD. We further demonstrate that NAS of Ig-μ minigene transcripts is not PTC specific. This finding, together with our results that contradict the previously reported frame dependence of TCR-β NAS, challenges the idea that cells might possess mechanisms that would allow regulation of splice site selection in response to premature termination of the ORF.