The Paraneoplastic Antigen Nova-1 is a Neuron Specific and Sequence Specific RNA Binding Protein

Author

Ronald J. Buckanovich

Date of Award

1996

Document Type

Thesis

Degree Name

Doctor of Philosophy (PhD)

RU Laboratory

Darnell Robert Laboratory

Keywords

Nova-1, RNA binding protein, alternative splicing, paraneoplastic opsoclonus-myoclonus ataxia (POMA), glycine receptor α2A (glyRα2A), KH motif

Abstract

We have identified a cDNA encoding a protein that is recognized by sera from (7/7) patients with paraneoplastic opsoclonus-myoclonus ataxia (POMA). The gene, Nova-1, encodes a novel nuclear RNA binding protein (RBP) that is extremely highly conserved (99% amino acid identity) between human and mouse. Nova-1 is normally expressed only within neurons of the central nervous system (CNS). Both Nova-1 protein and RNA are restricted to a subset of CNS neurons which correlate with the specific neurologic symptoms seen in POMA. Nova-1 contains three KH type RNA binding motifs, and binds RNA in vitro via these KH motifs. Mutations within the core of the KH motif abolish specific RNA binding. Utilizing RNA selection, we have identified an in vitro RNA ligand for Nova-1 consisting of three UCAUY repeats in the loop of a stem loop structure. We have identified a similar sequence upstream of the alternatively spliced exons (3A and 3B) of the Glycine receptor α2A (glyRα2A), and downstream of the alternatively spliced exon H of Nova-1, and demonstrate that Nova-1 binds these RNAs with high affinity (Kd ~2 nM). We have also shown that Nova-1 binds both the glyRα2A and Nova-1 pre-mRNAs in vivo. Nova-1 binding to the target sequence in the glyRα2A RNA in tissue culture cells stimulates splicing to exon 3A suggesting that Nova-1 functions in regulating alternative splicing in mouse brain. Moreover, the demonstration that paraneoplastic antibodies can prevent Nova-1 from binding to glyRα2A RNA in vitro suggests a potential mechanism for the neurologic symptoms of POMA.

Comments

A thesis presented to the faculty of The Rockefeller University in partial fulfillment of the requirements for the degree of Doctor of Philosophy

Recommended Citation

Buckanovich, Ronald J., "The Paraneoplastic Antigen Nova-1 is a Neuron Specific and Sequence Specific RNA Binding Protein" (1996). Student Theses and Dissertations. 338.
https://digitalcommons.rockefeller.edu/student_theses_and_dissertations/338