Role of the single nucleotide polymorphism rs7903146 of TCF7L2 in inducing nonsense-mediated decay.
BACKGROUND
The single nucleotide polymorphism (SNP) rs7903146 (C/T), located in intron 4 of the transcription factor 7-like 2 gene (TCF7L2), has been associated with an increased risk of developing Type 2 Diabetes, although the molecular mechanism remain elusive. The TCF7L2 gene is alternatively spliced but an association between genotype and splice variants has not been shown convincingly. We hypothesized that a yet unknown extra exon, containing either the C or T genotype of the SNP rs7903146, could introduce a premature stop codon and consequently result in nonsense-mediated decay (NMD).


RESULTS
Running the sequences C and T of the SNP region in different servers we found that the two alleles could display differential recognition by splicing factors. The C variant showed the possible inclusion of an unknown exon. This unknown exon contained a stop codon and thus could induce NMD. We then determined that the splicing pattern in isolated mouse islets and MIN6 cells was similar to that in human pancreatic islets. Therefore, we used MIN6 cells to study the splicing of human intron 4: two mini-genes of intron 4 containing either the C/C genotype or the T/T genotype were transfected into MIN6 cells. Our constructs were spliced normally, excluding intron 4, but we did not observe the presence of an extra exon with either construct.


CONCLUSIONS
We found that an extra exon could theoretically exist, although we were not able to capture it in our model. A better model is needed to determine whether a theoretical extra exon can induce NMD.
Diabetes and Obesity Research Laboratory, Hospital Clinic, University of Barcelona, IDIBAPS, Rosselló 153, Barcelona, Spain.
Nathalie
N
Nicod
Diabetes and Obesity Research Laboratory, Hospital Clinic, University of Barcelona, IDIBAPS, Rosselló 153, Barcelona, Spain.
Marta
M
Pradas-Juni
Diabetes and Obesity Research Laboratory, Hospital Clinic, University of Barcelona, IDIBAPS, Rosselló 153, Barcelona, Spain.
Ramon
R
Gomis
Alternative splicing; Beta-cell; Islets; Nonsense-mediated decay; SNP rs7903146; TCF7L2; Type 2 diabetes