Opioid precursor protein isoform is targeted to the cell nuclei in the human brain
| العنوان: | Opioid precursor protein isoform is targeted to the cell nuclei in the human brain |
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| المؤلفون: | Kononenko, Olga, Bazov, Igor, 1973, Watanabe, Hiroyuki, Gerashchenko, Ganna, Dyachok, Oleg, Verbeek, Dineke S, Alkass, Kanar, Druid, Henrik, Andersson, Malin, Mulder, Jan, Svenningsen, Åsa Fex, Rajkowska, Grazyna, Stockmeier, Craig A, Krishtal, Oleg, Yakovleva, Tatiana, Bakalkin, Georgy |
| المصدر: | Biochimica et Biophysica Acta - General Subjects. 1861(2):246-255 |
| مصطلحات موضوعية: | Alternative splicing, Human brain, Neuropeptide precursor protein, Nuclear localization, Prodynorphin |
| الوصف: | BACKGROUND: Neuropeptide precursors are traditionally viewed as proteins giving rise to small neuropeptide molecules. Prodynorphin (PDYN) is the precursor protein to dynorphins, endogenous ligands for the κ-opioid receptor. Alternative mRNA splicing of neuropeptide genes may regulate cell- and tissue-specific neuropeptide expression and produce novel protein isoforms. We here searched for novel PDYN mRNA and their protein product in the human brain.METHODS: Novel PDYN transcripts were identified using nested PCR amplification of oligo(dT) selected full-length capped mRNA. Gene expression was analyzed by qRT-PCR, PDYN protein by western blotting and confocal imaging, dynorphin peptides by radioimmunoassay. Neuronal nuclei were isolated using fluorescence-activated nuclei sorting (FANS) from postmortem human striatal tissue. Immunofluorescence staining and confocal microscopy was performed for human caudate nucleus.RESULTS: Two novel human PDYN mRNA splicing variants were identified. Expression of one of them was confined to the striatum where its levels constituted up to 30% of total PDYN mRNA. This transcript may be translated into ∆SP-PDYN protein lacking 13 N-terminal amino acids, a fragment of signal peptide (SP). ∆SP-PDYN was not processed to mature dynorphins and surprisingly, was targeted to the cell nuclei in a model cellular system. The endogenous PDYN protein was identified in the cell nuclei in human striatum by western blotting of isolated neuronal nuclei, and by confocal imaging.CONCLUSIONS AND GENERAL SIGNIFICANCE: High levels of alternatively spliced ∆SP-PDYN mRNA and nuclear localization of PDYN protein suggests a nuclear function for this isoform of the opioid peptide precursor in human striatum. |
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| URL الوصول: | https://urn.kb.se/resolve?urn=urn:nbn:se:oru:diva-82996 https://doi.org/10.1016/j.bbagen.2016.11.002 |
| قاعدة البيانات: | SwePub |
| تدمد: | 03044165 18728006 |
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| DOI: | 10.1016/j.bbagen.2016.11.002 |