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PMID: 9154818 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The neuronal RNA binding protein Nova-1 recognizes specific RNA targets in vitro and in vivo.

Molecular and cellular biology ·Vol. 17 ·No. 6 ·1997-06-00 ·Pages 3194-201

Buckanovich RJ, Darnell RB

Abstract

Nova-1, an autoantigen in paraneoplastic opsoclonus myoclonus ataxia (POMA), a disorder associated with breast cancer and motor dysfunction, is a neuron-specific nuclear RNA binding protein. We have identified in vivo Nova-1 RNA ligands by combining affinity-elution-based RNA selection with protein-RNA immunoprecipitation. Starting with a pool of approximately 10(15) random 52-mer RNAs, we identified long stem-loop RNA ligands that bind to Nova-1 with high affinity (Kd of approximately 2 nM). The loop region of these RNAs harbors a approximately 15-bp pyrimidine-rich element [UCAU(N)(0-2)]3 which is essential for Nova-1 binding. Mutagenesis studies defined the third KH domain of Nova-1 and the [UCAU(N)(0-2)]3 element as necessary for in vitro binding. Consensus [UCAU (N)(0-2)], elements were identified in two neuronal pre-mRNAs, one encoding the inhibitory glycine receptor alpha2 (GlyR alpha2) and a second encoding Nova-1 itself. Nova-1 protein binds these RNAs with high affinity and specificity in vitro, and this binding can be blocked by POMA antisera. Moreover, both Nova-1 and GlyR alpha2 pre-mRNAs specifically coimmunoprecipitated with Nova-1 protein from brain extracts. Thus, Nova-1 functions as a sequence-specific nuclear RNA binding protein in vivo; disruption of the specific interaction between Nova-1 and GlyR alpha2 pre-mRNA may underlie the motor dysfunction seen in POMA.

MeSH Terms
Animals Antigens, Neoplasm/metabolism Ataxia/immunology Base Sequence Consensus Sequence In Vitro Techniques Mice Molecular Sequence Data Nerve Tissue Proteins/metabolism Neuro-Oncological Ventral Antigen Neurons/chemistry Nucleic Acid Conformation Paraneoplastic Syndromes/immunology Polymerase Chain Reaction RNA/metabolism RNA-Binding Proteins/metabolism Ribonucleoproteins/metabolism Structure-Activity Relationship
Chemicals
Antigens, Neoplasm Nerve Tissue Proteins Neuro-Oncological Ventral Antigen RNA-Binding Proteins Ribonucleoproteins RNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Buckanovich R J
Laboratory of Molecular Neuro-Oncology, The Rockefeller University, New York, New York 10021, USA.
Darnell R B
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.6.3194
Published
1997-06-00
Pages
3194-201
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232172
Subset
IM
Grants
NIGMS NIH HHS · 5T32 GM07739 · United States
NINDS NIH HHS · R01 NS34389 · United States
External Links
PubMed source DOI: 10.1128/MCB.17.6.3194 PMC Full Text View journal details
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