Reversion mutation of cDNA CA8-204 minigene construct produces a truncated functional peptide that regulates calcium release in vitro and produces profound analgesia in vivo
- 28 November 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Mammalian Genome
- Vol. 31 (9-12), 287-294
- https://doi.org/10.1007/s00335-020-09848-y
Abstract
Intracellular calcium is critical in orchestrating neuronal excitability and analgesia. Carbonic anhydrase-8 (CA8) regulates intracellular calcium signaling through allosteric inhibition of neuronal inositol trisphosphate receptor 1 (ITPR1) to produce profound analgesia. Recently, we reported the “G” allele at rs6471859 represents cis-eQTL regulating alternative splicing of a 1697 bp transcript (CA8-204G) with a retained intron, alternative polyadenylation site and a new stop codon producing a functional 26 kDa peptide with an extended exon 3. In this study we show the reversion mutation (G to C) at rs6471859 within the CA8-204G expression vector also produced a stable 1697 bp transcript (CA8-204C) coding for a smaller peptide (~ 22 kDa) containing only the first three CA8 exons. Surprisingly, this peptide inhibited ITPR1 (pITPR1) activation, ITPR1-mediated calcium release in vitro; and produced profound analgesia in vivo. This is the first report showing CA8-204C codes for a functional peptide sufficient to regulate calcium signaling and produce profound analgesia.Keywords
Funding Information
- National Institute of Neurological Disorders and Stroke (NS105880, 3R21NS105880-01S1)
- National Institute of Dental and Craniofacial Research (DE022903)
- DOD Counterdrug Technology Development Program Office (W81XWH-19-1-0525)
This publication has 15 references indexed in Scilit:
- Impact of human CA8 on thermal antinociception in relation to morphine equivalence in miceNeuroReport, 2017
- Alternative RNA splicing: contribution to pain and potential therapeutic strategyDrug Discovery Today, 2016
- Painful and painless channelopathiesThe Lancet Neurology, 2014
- Voltage-gated sodium channel mutations and painful neuropathy: Na(v)1.9 joins the family.Brain, 2014
- Cell-Based Splicing of MinigenesMethods in Molecular Biology, 2014
- Alternative splicing generates a truncated isoform of human TNFRSF11A (RANK) with an altered capacity to activate NF-κBGene, 2013
- Carbonic anhydrase related protein 8 mutation results in aberrant synaptic morphology and excitatory synaptic function in the cerebellumMolecular and Cellular Neuroscience, 2007
- Widespread mRNA polyadenylation events in introns indicate dynamic interplay between polyadenylation and splicingGenome Research, 2007
- TAT gene mutation analysis in three Palestinian kindreds with oculocutaneous tyrosinaemia type II; characterization of a silent exonic transversion that causes complete missplicing by exon 11 skippingJournal of Inherited Metabolic Disease, 2006
- Carbonic anhydrase-related protein is a novel binding protein for inositol 1,4,5-trisphosphate receptor type 1Biochemical Journal, 2003