A P2RX7 single nucleotide polymorphism haplotype promotes exon 7 and 8 skipping and disrupts receptor function

Skarratt, Kristen K., Gu, Ben J., Lovelace, Michael D., Milligan, Carol J., Stokes, Leanne ORCID: https://orcid.org/0000-0003-4013-6781, Glover, Rachel, Petrou, Steven, Wiley, James S. and Fuller, Stephen J. (2020) A P2RX7 single nucleotide polymorphism haplotype promotes exon 7 and 8 skipping and disrupts receptor function. The FASEB Journal, 34 (3). pp. 3884-3901. ISSN 0892-6638

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Abstract

P2X7 is an ATP-gated membrane ion channel that is expressed by multiple cell types. Brief exposure to ATP induces the opening of a nonselective cation channel; while repeated or prolonged exposure induces formation of a transmembrane pore. This process may be partially regulated by alternative splicing of full-length P2RX7A pre-mRNA, producing isoforms that delete or retain functional domains. Here, we report cloning and expression of a novel P2RX7 splice variant, P2RX7L, that is, characterized by skipping of exons 7 and 8. In HEK 293 cells, expression of P2RX7L produces a protein isoform, P2X7L, that forms a heteromer with P2X7A. A haplotype defined by six single nucleotide polymorphisms (SNPs) (rs208307, rs208306, rs36144485, rs208308, rs208309, and rs373655596) promotes allele-specific alternative splicing, increasing mRNA levels of P2RX7L and another isoform, P2RX7E, which in addition has a truncated C-terminus. Skipping of exons 7 and 8 is predicted to delete critical amino acids in the ATP-binding site. P2X7L-transfected HEK 293 cells have phagocytic but not channel, pore, or membrane-blebbing function, and double-transfected P2X7L and P2X7A cells have reduced pore function. Heteromeric receptor complexes of P2X7A and P2X7L are predicted to have reduced numbers of ATP-binding sites, which potentially alters receptor function compared to homomeric P2X7A complexes.

Item Type: Article
Additional Information: © 2020 Federation of American Societies for Experimental Biology.
Uncontrolled Keywords: p2x7,alternative splicing,heteromer,single nucleotide polymorphism,allele,cancer-cells,activation,c-terminus,quantitative method,pore formation,atp-binding,p2x(7) receptor,p2x7 receptor,ion-channel,genetics,molecular biology,biochemistry,biotechnology,sdg 3 - good health and well-being ,/dk/atira/pure/subjectarea/asjc/1300/1311
Faculty \ School: Faculty of Science > School of Pharmacy
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Depositing User: LivePure Connector
Date Deposited: 17 Apr 2020 00:50
Last Modified: 22 Oct 2022 05:53
URI: https://ueaeprints.uea.ac.uk/id/eprint/74771
DOI: 10.1096/fj.201901198RR

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