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https://hdl.handle.net/2440/34837
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Type: | Journal article |
Title: | Nova regulates GABA(A) receptor gamma 2 alternative splicing via a distal downstream UCAU-rich intronic splicing enhancer |
Author: | Dredge, B. Darnell, R. |
Citation: | Molecular and Cellular Biology, 2003; 23(13):4687-4700 |
Publisher: | Amer Soc Microbiology |
Issue Date: | 2003 |
ISSN: | 0270-7306 1098-5549 |
Statement of Responsibility: | B. Kate Dredge and Robert B. Darnell |
Abstract: | Nova is a neuron-specific RNA binding protein targeted in patients with the autoimmune disorder paraneoplastic opsoclonus-myoclonus ataxia, which is characterized by failure of inhibition of brainstem and spinal motor systems. Here, we have biochemically confirmed the observation that splicing regulation of the inhibitory GABA(A) receptor gamma2 (GABA(A)Rgamma2) subunit pre-mRNA exon E9 is disrupted in mice lacking Nova-1. To elucidate the mechanism by which Nova-1 regulates GABA(A)Rgamma2 alternative splicing, we systematically screened minigenes derived from the GABA(A)Rgamma2 and human beta-globin genes for their ability to support Nova-dependent splicing in transient transfection assays. These studies demonstrate that Nova-1 acts directly on GABA(A)Rgamma2 pre-mRNA to regulate E9 splicing and identify an intronic region that is necessary and sufficient for Nova-dependent enhancement of exon inclusion, which we term the NISE (Nova-dependent intronic splicing enhancer) element. The NISE element (located 80 nucleotides upstream of the splice acceptor site of the downstream exon E10) is composed of repeats of the sequence YCAY, consistent with previous studies of the mechanism by which Nova binds RNA. Mutation of these repeats abolishes binding of Nova-1 to the RNA in vitro and Nova-dependent splicing regulation in vivo. These data provide a molecular basis for understanding Nova regulation of GABA(A)Rgamma2 alternative splicing and suggest that general dysregulation of Nova's splicing enhancer function may underlie the neurologic defects seen in Nova's absence. |
Keywords: | Cell Line Animals Mice, Transgenic Humans Mice Globins RNA-Binding Proteins Receptors, GABA-A Nerve Tissue Proteins Recombinant Fusion Proteins DNA, Complementary RNA, Messenger Antigens, Neoplasm Collodion Blotting, Western Transfection Reverse Transcriptase Polymerase Chain Reaction Gene Expression Regulation Alternative Splicing Base Sequence Protein Binding Sequence Homology, Nucleic Acid Dose-Response Relationship, Drug Mutation Introns Exons Plasmids Models, Genetic Molecular Sequence Data Enhancer Elements, Genetic Neuro-Oncological Ventral Antigen |
Description: | Copyright © 2003, American Society for Microbiology |
DOI: | 10.1128/MCB.23.13.4687-4700.2003 |
Published version: | http://dx.doi.org/10.1128/mcb.23.13.4687-4700.2003 |
Appears in Collections: | Aurora harvest 6 Molecular and Biomedical Science publications |
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