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Expression and translation of the COX-1b gene in human cells – no evidence of generation of COX-1b protein

  • Christina Reinauer , Petra Censarek , Gernot Kaber , Artur-Aron Weber , Gerhard Steger , Thorsten Klamp and Karsten Schrör EMAIL logo
Published/Copyright: January 28, 2013

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.


Corresponding author: Karsten Schrör, Institute for Pharmacology and Clinical Pharmacology, Heinrich-Heine-University, D-40225 Düsseldorf, Germany

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).

References

Bendtsen, J.D., Nielsen, H., von Heijne, G., and Brunak, S. (2004). Improved prediction of signal peptides: SignalP 3.0. J. Mol. Biol. 340, 783–795.10.1016/j.jmb.2004.05.028Search in Google Scholar

Censarek, P., Freidel, K., Udelhoven, M., Ku, S.J., Hohlfeld, T., Meyer-Kirchrath, J., Schrör, K., and Weber, A.A. (2004). Cyclooxygenase COX-2a, a novel COX-2 mRNA variant, in platelets from patients after coronary artery bypass grafting. Thromb. Haemost. 92, 925–928.10.1160/TH04-05-0302Search in Google Scholar

Censarek, P., Freidel, K., Hohlfeld, T., Schrör, K., and Weber, A.A. (2006). Human cyclooxygenase-1b is not the elusive target of acetaminophen. Eur. J. Pharmacol. 551, 50–53.10.1016/j.ejphar.2006.08.079Search in Google Scholar

Chandrasekharan, N.V., Dai, H., Roos, K.L., Evanson, N.K., Tomsik, J., Elton, T.S., and Simmons, D.L. (2002). COX-3, a cyclooxygenase-1 variant inhibited by acetaminophen and other analgesic/antipyretic drugs: cloning, structure, and expression. Proc. Natl. Acad. Sci. USA 99, 13926–13931.10.1073/pnas.162468699Search in Google Scholar

Cui, J.G., Kuroda, H., Chandrasekharan, N.V., Pelaez, R.P., Simmons, D.L., Bazan, N.G., and Lukiw, W.J. (2004). Cyclooxygenase-3 gene expression in Alzheimer hippocampus and in stressed human neural cells. Neurochem. Res. 29, 1731–1737.10.1023/B:NERE.0000035809.70905.8aSearch in Google Scholar

Dinchuk, J.E., Liu, R.Q., and Trzaskos, J.M. (2003). COX-3: in the wrong frame in mind. Immunol. Lett. 86, 121.10.1016/S0165-2478(02)00268-7Search in Google Scholar

Emanuelsson, O., Brunak, S., von Heijne, G., and Nielsen, H. (2007). Locating proteins in the cell using TargetP, SignalP and related tools. Nat. Protoc. 2, 953–971.10.1038/nprot.2007.131Search in Google Scholar PubMed

Graham, G.G. and Scott, K.F. (2005). Mechanism of action of paracetamol. Am. J. Ther. 12, 46–55.10.1097/00045391-200501000-00008Search in Google Scholar PubMed

Ivanov, I.P., Gesteland, R.F., Matsufuji, S., and Atkins, J.F. (1998). Programmed frameshifting in the synthesis of mammalian antizyme is +1 in mammals, predominantly +1 in fission yeast, but -2 in budding yeast. RNA 4, 1230–1238.10.1017/S1355838298980864Search in Google Scholar PubMed PubMed Central

Kam, P.C.A. and So, A. (2009). COX-3: uncertainties and controversies. Curr. Anaesth. Crit. Care 20, 50–53.10.1016/j.cacc.2008.11.003Search in Google Scholar

Kis, B., Snipes, J.A., Isse, T., Nagy, K., and Busija, D.W. (2003). Putative cyclooxygenase-3 expression in rat brain cells. J. Cereb. Blood Flow Metab. 23, 1287–1292.10.1097/01.WCB.0000090681.07515.81Search in Google Scholar PubMed

Kis, B., Snipes, J.A., Gaspar, T., Lenzser, G., Tulbert, C.D., and Busija, D.W. (2006). Cloning of cyclooxygenase-1b (putative COX-3) in mouse. Inflamm. Res. 55, 274–278.10.1007/s00011-006-0083-zSearch in Google Scholar PubMed

Klamp, T., Sahin, U., Kyewski, B., Schwendemann, J., Dhaene, K., and Türeci, O. (2006). Expression profiling of autoimmune regulator AIRE mRNA in a comprehensive set of human normal and neoplastic tissues. Immunol. Lett. 106, 172–179.10.1016/j.imlet.2006.06.006Search in Google Scholar PubMed

Moon, S., Byun, Y., Kim, H.J., Jeong, S., and Han, K. (2004). Predicting genes expressed via -1 and +1 frameshifts. Nucleic. Acid. Res. 32, 4884–4892.10.1093/nar/gkh829Search in Google Scholar PubMed PubMed Central

Nurmi, J.T., Puolakkainen, P.A., and Rautonen, N.E. (2005). Intron 1 retaining cyclooxygenase 1 splice variant is induced by osmotic stress in human intestinal epithelial cells. Prostaglandins Leukot. Essent. Fatty Acids 73, 343–350.10.1016/j.plefa.2005.07.004Search in Google Scholar PubMed

Qin, N., Zhang, S.P., Reitz, T.L., Mei, J.M., and Flores, C.M. (2005). Cloning, expression, and functional characterization of human cyclooxygenase-1 splicing variants: evidence for intron 1 retention. J. Pharmacol. Exp. Ther. 315, 1298–1305.10.1124/jpet.105.090944Search in Google Scholar PubMed

Reeder, J. and Giegerich, R. (2004). Design, implementation and evaluation of a practical pseudo-knot algorithm based on thermodynamics. BMC Bioinformatics. 5, 104.10.1186/1471-2105-5-104Search in Google Scholar PubMed PubMed Central

Schaefer, H., Chervet, J.P., Bunse, C., Joppich, C., Meyer, H.E., and Marcus, K.A. (2004). Peptide preconcentration approach for nano-high-performance liquid chromatography to diminish memory effects. Proteomics 4, 2541–2544.10.1002/pmic.200300801Search in Google Scholar PubMed

Schwab, J.M., Beiter, T., Linder, J.U., Laufer, S., Schulz, J.E., Meyermann, R., and Schluesener, H.J. (2003). COX-3 – a virtual pain target in humans? FASEB J. 17, 2174–2175.Search in Google Scholar

Simmons, D.L., Botting, R.M., and Hla, T. (2004). Cyclooxygenase isozymes: the biology of prostaglandin synthesis and inhibition. Pharmacol. Rev. 56, 387–437.10.1124/pr.56.3.3Search in Google Scholar PubMed

Snipes, J.A., Kis, B., Shelness, G.S., Hewett, J.A., and Busija, D.W. (2005). Cloning and characterization of cyclooxygenase-1b (putative cyclooxygenase-3) in rat. J. Pharmacol. Exp. Ther. 313, 668–676.10.1124/jpet.104.079533Search in Google Scholar PubMed

Willoughby, D.A., Moore, A.R., and Colville-Nash, P.R. (2000). COX-1, COX-2, and COX-3 and the future treatment of chronic inflammatory disease. Lancet 355, 646–648.10.1016/S0140-6736(99)12031-2Search in Google Scholar

Wills, N.M. and Atkins, J.F. (2006). The potential role of ribosomal frameshifting in generating aberrant proteins implicated in neurodegenerative diseases. RNA 12, 1149–1153.10.1261/rna.84406Search in Google Scholar PubMed PubMed Central

Received: 2012-10-19
Accepted: 2013-1-14
Published Online: 2013-01-28
Published in Print: 2013-06-01

©2013 by Walter de Gruyter Berlin Boston

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