Abstract
We have combined genetic and biochemical approaches to analyze the function of the RNA-binding protein Nova-1, the paraneoplastic opsoclonus-myoclonus ataxia (POMA) antigen. Nova-1 null mice die postnatally from a motor deficit associated with apoptotic death of spinal and brainstem neurons. Nova-1 null mice show specific splicing defects in two inhibitory receptor pre-mRNAs, glycine alpha2 exon 3A (GlyRalpha2 E3A) and GABA(A) exon gamma2L. Nova protein in brain extracts specifically bound to a previously identified GlyRalpha2 intronic (UCAUY)3 Nova target sequence, and Nova-1 acted directly on this element to increase E3A splicing in cotransfection assays. We conclude that Nova-1 binds RNA in a sequence-specific manner to regulate neuronal pre-mRNA alternative splicing; the defect in splicing in Nova-1 null mice provides a model for understanding the motor dysfunction in POMA.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Alternative Splicing / physiology*
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Animals
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Antigens, Neoplasm*
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Apoptosis / genetics
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Brain Chemistry / genetics
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Brain Stem / cytology
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Brain Stem / embryology
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Cell Survival / genetics
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Exons / genetics
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Gene Deletion
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Gene Expression Regulation, Developmental
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Genotype
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In Situ Nick-End Labeling
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Motor Neurons / chemistry
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Motor Neurons / cytology*
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Motor Neurons / physiology*
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Nerve Tissue Proteins*
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Neuro-Oncological Ventral Antigen
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Protein Binding / genetics
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RNA Precursors / genetics
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RNA-Binding Proteins / genetics*
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RNA-Binding Proteins / metabolism
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Receptors, GABA-A / genetics
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Receptors, GABA-A / metabolism
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Receptors, Glycine / genetics
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Receptors, Glycine / metabolism
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Ribonucleoproteins / genetics*
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Ribonucleoproteins / metabolism
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Spinal Cord / cytology
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Spinal Cord / embryology
Substances
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Antigens, Neoplasm
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Nerve Tissue Proteins
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Neuro-Oncological Ventral Antigen
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RNA Precursors
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RNA-Binding Proteins
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Receptors, GABA-A
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Receptors, Glycine
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Ribonucleoproteins