Protein-RNA and protein-protein recognition by dual KH1/2 domains of the neuronal splicing factor Nova-1.

Abstract

Nova onconeural antigens are neuron-specific RNA-binding proteins implicated in paraneoplastic opsoclonus-myoclonus-ataxia (POMA) syndrome. Nova harbors three K-homology (KH) motifs implicated in alternate splicing regulation of genes involved in inhibitory synaptic transmission. We report the crystal structure of the first two KH domains (KH1/2) of Nova-1 bound to an in vitro selected RNA hairpin, containing a UCAG-UCAC high-affinity binding site. Sequence-specific intermolecular contacts in the complex involve KH1 and the second UCAC repeat, with the RNA scaffold buttressed by interactions between repeats. Whereas the canonical RNA-binding surface of KH2 in the above complex engages in protein-protein interactions in the crystalline state, the individual KH2 domain can sequence-specifically target the UCAC RNA element in solution. The observed antiparallel alignment of KH1 and KH2 domains in the crystal structure of the complex generates a scaffold that could facilitate target pre-mRNA looping on Nova binding, thereby potentially explaining Nova's functional role in splicing regulation.

• Authors: Marianna Teplova, Lucy Malinina, Jennifer C Darnell, Jikui Song, Min Lu, Ruben Abagyan, Kiran Musunuru, Alexei Teplov, Stephen K Burley, Robert B Darnell, Dinshaw J Patel
• Journal: Structure (London, England : 1993) (Structure) Vol. 19 Issue 7 Pg. 930-44 (Jul 13 2011) ISSN: 1878-4186 [Electronic] United States
• PMID: 21742260 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2011 Elsevier Ltd. All rights reserved.

Chemical References

• Antigens, Neoplasm
• Nerve Tissue Proteins
• Neuro-Oncological Ventral Antigen
• RNA Precursors
• RNA, Small Interfering
• RNA-Binding Proteins
• Solutions

Topics

• Alternative Splicing
• Amino Acid Sequence
• Antigens, Neoplasm (chemistry, genetics, metabolism)
• Base Sequence
• Binding Sites
• Crystallization
• Crystallography, X-Ray
• Electrophoretic Mobility Shift Assay
• Humans
• Magnetic Resonance Spectroscopy
• Models, Molecular
• Molecular Sequence Data
• Nerve Tissue Proteins (chemistry, genetics, metabolism)
• Neuro-Oncological Ventral Antigen
• Neurons (cytology, metabolism)
• Opsoclonus-Myoclonus Syndrome (metabolism, physiopathology)
• Protein Binding
• Protein Structure, Tertiary
• RNA Precursors (chemistry, metabolism)
• RNA, Small Interfering (chemistry, metabolism)
• RNA-Binding Proteins (chemistry, genetics, metabolism)
• Sequence Homology, Amino Acid
• Solutions (chemistry, metabolism)
• Synaptic Transmission (physiology)
• Syndrome