RNA‐editing of the 5‐HT2C receptor alters agonist‐receptor‐effector coupling specificity

Department of Pharmacology, Mail Code 7764, University of Texas Health Science Center, San Antonio, Texas, TX 78229-3900, USA.
British Journal of Pharmacology (Impact Factor: 4.84). 10/2001; 134(2):386-92. DOI: 10.1038/sj.bjp.0704255
Source: PubMed

ABSTRACT 1. The serotonin(2C) (5-HT(2C)) receptor couples to both phospholipase C (PLC)-inositol phosphate (IP) and phospholipase A(2) (PLA(2))-arachidonic acid (AA) signalling cascades. Agonists can differentially activate these effectors (i.e. agonist-directed trafficking of receptor stimulus) perhaps due to agonist-specific receptor conformations which differentially couple to/activate transducer molecules (e.g. G proteins). Since editing of RNA transcripts of the human 5-HT(2C) receptor leads to substitution of amino acids at positions 156, 158 and 160 of the putative second intracellular loop, a region important for G protein coupling, we examined the capacity of agonists to activate both the PLC-IP and PLA(2)-AA pathways in CHO cells stably expressing two major, fully RNA-edited isoforms (5-HT(2C-VSV), 5-HT(2C-VGV)) of the h5-HT(2C) receptor. 2. 5-HT increased AA release and IP accumulation in both 5-HT(2C-VSV) and 5-HT(2C-VGV) expressing cells. As expected, the potency of 5-HT for both RNA-edited isoforms for both responses was 10 fold lower relative to that of the non-edited receptor (5-HT(2C-INI)) when receptors were expressed at similar levels. 3. Consistent with our previous report, the efficacy order of two 5-HT receptor agonists (TFMPP and bufotenin) was reversed for AA release and IP accumulation at the non-edited receptor thus demonstrating agonist trafficking of receptor stimulus. However, with the RNA-edited receptor isoforms there was no difference in the relative efficacies of TFMPP or bufotenin for AA release and IP accumulation suggesting that the capacity for 5-HT(2C) agonists to traffic receptor stimulus is lost as a result of RNA editing. 4. These results suggest an important role for the second intracellular loop in transmitting agonist-specific information to signalling molecules.

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Available from: Kelly A Berg, Aug 18, 2014
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    • "Ribosomes decode inosine as guanosine, and editing events within open reading frames lead to production of edited isoforms of ion channel subunits and other proteins. Production of edited subunit isoforms affects the pharmacological properties of channels and receptors such as AMPA receptors, potassium Kv1.1 channels and serotonin 5HT2c receptors (3–6). Various neurological disorders including Motor Neuron Disease, depression and epilepsy have been linked to defects in RNA editing, particularly to ADAR2 and to the transcripts that it edits (7–11). "
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    ABSTRACT: RNA editing by deamination of specific adenosine bases to inosines during pre-mRNA processing generates edited isoforms of proteins. Recoding RNA editing is more widespread in Drosophila than in vertebrates. Editing levels rise strongly at metamorphosis, and Adar(5G1) null mutant flies lack editing events in hundreds of CNS transcripts; mutant flies have reduced viability, severely defective locomotion and age-dependent neurodegeneration. On the other hand, overexpressing an adult dADAR isoform with high enzymatic activity ubiquitously during larval and pupal stages is lethal. Advantage was taken of this to screen for genetic modifiers; Adar overexpression lethality is rescued by reduced dosage of the Rdl (Resistant to dieldrin), gene encoding a subunit of inhibitory GABA receptors. Reduced dosage of the Gad1 gene encoding the GABA synthetase also rescues Adar overexpression lethality. Drosophila Adar(5G1) mutant phenotypes are ameliorated by feeding GABA modulators. We demonstrate that neuronal excitability is linked to dADAR expression levels in individual neurons; Adar-overexpressing larval motor neurons show reduced excitability whereas Adar(5G1) null mutant or targeted Adar knockdown motor neurons exhibit increased excitability. GABA inhibitory signalling is impaired in human epileptic and autistic conditions, and vertebrate ADARs may have a relevant evolutionarily conserved control over neuronal excitability.
    Nucleic Acids Research 10/2013; 42(2). DOI:10.1093/nar/gkt909 · 9.11 Impact Factor
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    • "The E, C, and D editing sites on premRNA overlap with the targeted region. Posttranscriptional A to I editing has been reported to decrease the efficiency of G-protein coupling and therefore generates 5HT2c receptor variants with reduced activity (Berg et al. 2001; Vitali et al. 2005). Hence, perfect base complementarities displayed by the antisense element of Snord115 RNA to the alternatively spliced and posttranscriptionally edited exon of 5HT2c premRNA suggest a tempting model for regulation of serotonin receptor biogenesis by snoRNAs (Cavaille et al. 2000). "
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    ABSTRACT: The evolution of new genes can ensue through either gene duplication and the neofunctionalization of one of the copies or the formation of a de novo gene from hitherto nonfunctional, neutrally evolving intergenic or intronic genomic sequences. Only very rarely are entire genes created de novo. Mostly, nonfunctional sequences are coopted as novel parts of existing genes, such as in the process of exonization whereby introns become exons through changes in splicing. Here, we report a case in which a novel non-protein coding RNA evolved by intron-sequence recruitment into their structures. cDNAs derived from rat brain small RNAs, revealed a novel snoRNA originating from one of the Snord115 copies in the rat Prader-Willi-Syndrome locus. We suggest that a single point substitution in the Snord115 region led to the expression of a longer snoRNA variant, designated as L-Snord115. Cell culture and footprinting experiments confirmed that a single nucleotide substitution at Snord115 position 67 destabilized the kink-turn (K-turn) motif within the canonical snoRNA, while distal intronic sequences provided an alternate D-box region. The exapted sequence displays putative base pairing to 28S rRNA and mRNA targets.
    Genome Biology and Evolution 10/2013; 5(11). DOI:10.1093/gbe/evt155 · 4.23 Impact Factor
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    • "Late-emerging vacuous chewing movements that occur weeks to months after treatment initiation can persist for months after drug withdrawal, and these vacuous chewing movements have been considered to model tardive dyskinesias (Waddington 1990). the precise mechanisms underlying vacuous chewing movements after long-term antipsychotic treatment are likely related to the primary mechanism of action of all antipsychotic drugs, namely blockade at the D 2 receptor (Wadenberg et al. 2001). the 5-Ht system and notably 5-Ht 2C receptor seem to be involved in the production of these debilitating motor side effects (Kostrzewa et al. 2007; Richtand et al. 2008). "
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    ABSTRACT: Serotonin2C (5-HT2C) receptors are expressed in the basal ganglia, a group of subcortical structures involved in the control of motor behaviour, mood and cognition. These receptors are mediating the effects of 5-HT throughout different brain areas via projections originating from midbrain raphe nuclei. A growing interest has been focusing on the function of 5-HT2C receptors in the basal ganglia because they may be involved in various diseases of basal ganglia function notably those associated with chronic impairment of dopaminergic transmission. 5-HT2C receptors act on numerous types of neurons in the basal ganglia, including dopaminergic, GABAergic, glutamatergic or cholinergic cells. Perhaps inherent to their peculiar molecular properties, the modality of controls exerted by 5-HT2C receptors over these cell populations can be phasic, tonic (dependent on the 5-HT tone) or constitutive (a spontaneous activity without the presence of the ligand). These controls are functionally organized in the basal ganglia: they are mainly localized in the input structures and preferentially distributed in the limbic/associative territories of the basal ganglia. The nature of these controls is modified in neuropsychiatric conditions such as Parkinson's disease, tardive dyskinesia or addiction. Most of the available data indicate that the function of 5-HT2C receptor is enhanced in cases of chronic alterations of dopamine neurotransmission. The review illustrates that 5-HT2C receptors play a role in maintaining continuous controls over the basal ganglia via multiple diverse actions. We will discuss their interest for treatments aimed at ameliorating current pharmacotherapies in schizophrenia, Parkinson's disease or drugs abuse.
    Experimental Brain Research 04/2013; 230(4). DOI:10.1007/s00221-013-3508-2 · 2.04 Impact Factor
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