CLIP identifies Nova-regulated RNA networks in the brain

J Ule, KB Jensen, M Ruggiu, A Mele, A Ule, RB Darnell - Science, 2003 - science.org
Science, 2003science.org
Nova proteins are neuron-specific antigens targeted in paraneoplastic opsoclonus
myoclonus ataxia (POMA), an autoimmune neurologic disease characterized by abnormal
motor inhibition. Nova proteins regulate neuronal pre-messenger RNA splicing by directly
binding to RNA. To identify Nova RNA targets, we developed a method to purify protein-RNA
complexes from mouse brain with the use of ultraviolet cross-linking and
immunoprecipitation (CLIP). Thirty-four transcripts were identified multiple times by Nova …
Nova proteins are neuron-specific antigens targeted in paraneoplastic opsoclonus myoclonus ataxia (POMA), an autoimmune neurologic disease characterized by abnormal motor inhibition.Nova proteins regulate neuronal pre-messenger RNA splicing by directly binding to RNA.To identify Nova RNA targets, we developed a method to purify protein-RNA complexes from mouse brain with the use of ultraviolet cross-linking and immunoprecipitation (CLIP).Thirty-four transcripts were identified multiple times by Nova CLIP.Three-quarters of these encode proteins that function at the neuronal synapse, and one-third are involved in neuronal inhibition.Splicing targets confirmed in Nova–/– mice include c-Jun N-terminal kinase 2, neogenin, and gephyrin; the latter encodes a protein that clusters inhibitory γ-aminobutyric acid and glycine receptors, two previously identified Nova splicing targets.Thus, CLIP reveals that Nova coordinately regulates a biologically coherent set of RNAs encoding multiple components of the inhibitory synapse, an observation that may relate to the cause of abnormal motor inhibition in POMA.
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