enzym ADAR2 Dotaz Zobrazit nápovědu
- Klíčová slova
- enzym ADAR2, perampanel,
- MeSH
- antikonvulziva terapeutické užití MeSH
- enzymy genetika MeSH
- epilepsie * etiologie farmakoterapie genetika MeSH
- lidé MeSH
- mutace MeSH
- vzácné nemoci etiologie farmakoterapie genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
ADAR RNA editing enzymes are high-affinity dsRNA-binding proteins that deaminate adenosines to inosines in pre-mRNA hairpins and also exert editing-independent effects. We generated a Drosophila AdarE374A mutant strain encoding a catalytically inactive Adar with CRISPR/Cas9. We demonstrate that Adar adenosine deamination activity is necessary for normal locomotion and prevents age-dependent neurodegeneration. The catalytically inactive protein, when expressed at a higher than physiological level, can rescue neurodegeneration in Adar mutants, suggesting also editing-independent effects. Furthermore, loss of Adar RNA editing activity leads to innate immune induction, indicating that Drosophila Adar, despite being the homolog of mammalian ADAR2, also has functions similar to mammalian ADAR1. The innate immune induction in fly Adar mutants is suppressed by silencing of Dicer-2, which has a RNA helicase domain similar to MDA5 that senses unedited dsRNAs in mammalian Adar1 mutants. Our work demonstrates that the single Adar enzyme in Drosophila unexpectedly has dual functions.
- MeSH
- adenosindeaminasa chemie genetika MeSH
- adenosinmonofosfát metabolismus MeSH
- bodová mutace genetika MeSH
- degenerace nervu patologie MeSH
- Drosophila melanogaster genetika imunologie MeSH
- editace RNA genetika MeSH
- katalýza MeSH
- lokomoce MeSH
- messenger RNA genetika metabolismus MeSH
- mozek metabolismus MeSH
- přirozená imunita genetika MeSH
- proteinové domény MeSH
- proteiny Drosophily chemie genetika metabolismus MeSH
- regulace genové exprese MeSH
- ribonukleasa III metabolismus MeSH
- RNA-helikasy metabolismus MeSH
- stárnutí patologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
The RNA editing enzyme ADAR2 is essential for the recoding of brain transcripts. Impaired ADAR2 editing leads to early-onset epilepsy and premature death in a mouse model. Here, we report bi-allelic variants in ADARB1, the gene encoding ADAR2, in four unrelated individuals with microcephaly, intellectual disability, and epilepsy. In one individual, a homozygous variant in one of the double-stranded RNA-binding domains (dsRBDs) was identified. In the others, variants were situated in or around the deaminase domain. To evaluate the effects of these variants on ADAR2 enzymatic activity, we performed in vitro assays with recombinant proteins in HEK293T cells and ex vivo assays with fibroblasts derived from one of the individuals. We demonstrate that these ADAR2 variants lead to reduced editing activity on a known ADAR2 substrate. We also demonstrate that one variant leads to changes in splicing of ADARB1 transcript isoforms. These findings reinforce the importance of RNA editing in brain development and introduce ADARB1 as a genetic etiology in individuals with intellectual disability, microcephaly, and epilepsy.
- MeSH
- adenosindeaminasa genetika MeSH
- alely MeSH
- alternativní sestřih genetika MeSH
- dítě MeSH
- genetická predispozice k nemoci genetika MeSH
- genetická variace genetika MeSH
- HEK293 buňky MeSH
- lidé MeSH
- mentální retardace genetika MeSH
- mikrocefalie genetika MeSH
- předškolní dítě MeSH
- proteiny vázající RNA genetika MeSH
- sestřih RNA genetika MeSH
- záchvaty genetika MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
BACKGROUND: Adenosine-to-inosine RNA editing is a co-transcriptional/post-transcriptional modification of double-stranded RNA, catalysed by one of two active adenosine deaminases acting on RNA (ADARs), ADAR1 and ADAR2. ADARB1 encodes the enzyme ADAR2 that is highly expressed in the brain and essential to modulate the function of glutamate and serotonin receptors. Impaired ADAR2 editing causes early onset progressive epilepsy and premature death in mice. In humans, ADAR2 dysfunction has been very recently linked to a neurodevelopmental disorder with microcephaly and epilepsy in four unrelated subjects. METHODS: We studied three children from two consanguineous families with severe developmental and epileptic encephalopathy (DEE) through detailed physical and instrumental examinations. Exome sequencing (ES) was used to identify ADARB1 mutations as the underlying genetic cause and in vitro assays with transiently transfected cells were performed to ascertain the impact on ADAR2 enzymatic activity and splicing. RESULTS: All patients showed global developmental delay, intractable early infantile-onset seizures, microcephaly, severe-to-profound intellectual disability, axial hypotonia and progressive appendicular spasticity. ES revealed the novel missense c.1889G>A, p.(Arg630Gln) and deletion c.1245_1247+1 del, p.(Leu415PhefsTer14) variants in ADARB1 (NM_015833.4). The p.(Leu415PhefsTer14) variant leads to incorrect splicing resulting in frameshift with a premature stop codon and loss of enzyme function. In vitro RNA editing assays showed that the p.(Arg630Gln) variant resulted in a severe impairment of ADAR2 enzymatic activity. CONCLUSION: In conclusion, these data support the pathogenic role of biallelic ADARB1 variants as the cause of a distinctive form of DEE, reinforcing the importance of RNA editing in brain function and development.
- MeSH
- adenosindeaminasa genetika metabolismus MeSH
- alely MeSH
- dítě MeSH
- dvouvláknová RNA metabolismus MeSH
- editace RNA MeSH
- epilepsie enzymologie genetika MeSH
- HEK293 buňky MeSH
- lidé MeSH
- mutace MeSH
- nemoci mozku enzymologie genetika metabolismus MeSH
- neurovývojové poruchy enzymologie genetika MeSH
- pokrevní příbuzenství MeSH
- předškolní dítě MeSH
- proteiny vázající RNA genetika metabolismus MeSH
- rodokmen MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
ADAR RNA editing enzymes (adenosine deaminases acting on RNA) that convert adenosine bases to inosines were first identified biochemically 30 years ago. Since then, studies on ADARs in genetic model organisms, and evolutionary comparisons between them, continue to reveal a surprising range of pleiotropic biological effects of ADARs. This review focuses on Drosophila melanogaster, which has a single Adar gene encoding a homolog of vertebrate ADAR2 that site-specifically edits hundreds of transcripts to change individual codons in ion channel subunits and membrane and cytoskeletal proteins. Drosophila ADAR is involved in the control of neuronal excitability and neurodegeneration and, intriguingly, in the control of neuronal plasticity and sleep. Drosophila ADAR also interacts strongly with RNA interference, a key antiviral defense mechanism in invertebrates. Recent crystal structures of human ADAR2 deaminase domain-RNA complexes help to interpret available information on Drosophila ADAR isoforms and on the evolution of ADARs from tRNA deaminase ADAT proteins. ADAR RNA editing is a paradigm for the now rapidly expanding range of RNA modifications in mRNAs and ncRNAs. Even with recent progress, much remains to be understood about these groundbreaking ADAR RNA modification systems.
- MeSH
- adenosindeaminasa chemie genetika metabolismus MeSH
- Drosophila melanogaster genetika metabolismus MeSH
- editace RNA * MeSH
- exprese genu MeSH
- interakční proteinové domény a motivy MeSH
- izoenzymy MeSH
- lidé MeSH
- messenger RNA genetika MeSH
- molekulární evoluce MeSH
- nervový systém metabolismus MeSH
- obratlovci MeSH
- proteiny Drosophily genetika metabolismus MeSH
- proteiny vázající RNA genetika metabolismus MeSH
- RNA interference MeSH
- substrátová specifita MeSH
- vazba proteinů MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH