title: 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: Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
authors: Marianna
authors: M
authors: Teplova
authors: Lucy
authors: L
authors: Malinina
authors: Jennifer C
authors: JC
authors: Darnell
authors: Jikui
authors: J
authors: Song
authors: Min
authors: M
authors: Lu
authors: Ruben
authors: R
authors: Abagyan
authors: Kiran
authors: K
authors: Musunuru
authors: Alexei
authors: A
authors: Teplov
authors: Stephen K
authors: SK
authors: Burley
authors: Robert B
authors: RB
authors: Darnell
authors: Dinshaw J
authors: DJ
authors: Patel
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D003460:Crystallization; D018360:Crystallography, X-Ray; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000090682:Neuro-Oncological Ventral Antigen; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D053578:Opsoclonus-Myoclonus Syndrome / Q000378:metabolism* / Q000503:physiopathology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012322:RNA Precursors / Q000737:chemistry / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000737:chemistry / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012996:Solutions / Q000737:chemistry / Q000378:metabolism; D009435:Synaptic Transmission / Q000502:physiology*; D013577:Syndrome
chemical_list: D000951:Antigens, Neoplasm; D009419:Nerve Tissue Proteins; D000090682:Neuro-Oncological Ventral Antigen; D012322:RNA Precursors; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D012996:Solutions