Expression and translation of the COX-1b gene in human cells – no evidence of generation of COX-1b protein
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Christina Reinauer
, Petra Censarek , Gernot Kaber , Artur-Aron Weber , Gerhard Steger , Thorsten Klamp and Karsten Schrör
Abstract
Cyclooxygenase 1b (COX-1b) is a splice variant of COX-1, containing a retained intron 1 within the signal peptide sequence. COX-1b mRNA is found in many species, but the existence of a functionally active protein, which is possibly related to different species-dependent lengths of intron 1, is controversially discussed. The human intron 1 comprises 94 bp, and the resulting frameshift at the intron 1-exon 2 junction creates a premature stop codon. Nevertheless, full-length human COX-1b protein expression, including translated intron 1 and the signal peptide, has been reported and was explained by a frameshift repair. In this study, the fate of COX-1b mRNA in a human overexpression system is analyzed. Independent of the hypothetical frameshift repair mechanism, the splicing of the COX-1b intron 1, resulting in COX-1 mRNA and removal of the signal peptide during protein maturation, with subsequent generation of a COX-1 protein is demonstrated.
The authors thank Kerstin Freidel and Beate Weyrauther for their excellent technical assistance. This study was approved by the local ethical review board (Ethikkommission der Ärztekammer des Landes Rheinland-Pfalz). The work was supported by the Deutsche Forschungsgemeinschaft (WE 2355/2-2) and the Forschungsgruppe Herz-Kreislauf e.V. (Düsseldorf).
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