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PMID: 10829067 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A brain-enriched polypyrimidine tract-binding protein antagonizes the ability of Nova to regulate neuron-specific alternative splicing.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 12 ·2000-06-06 ·Pages 6350-5

Polydorides AD, Okano HJ, Yang YY, Stefani G, Darnell RB

Abstract

The Nova paraneoplastic antigens are neuron-specific RNA binding proteins that participate in the control of alternative splicing. We have used the yeast two-hybrid system to isolate Nova interacting proteins and identify an RNA binding protein that is closely related to the polypyrimidine tract-binding protein (PTB). The expression of this protein, brPTB, is enriched in the brain, where it is expressed in glia and neurons. brPTB interacts with Nova proteins in cell lines and colocalizes with Nova within neuronal nuclei. We previously found that Nova binds to a pyrimidine-rich RNA element present upstream of an alternatively spliced exon, E3A, in glycine receptor alpha2 (GlyRalpha2) pre-mRNA, and this binding is implicated in Nova-dependent regulation of splicing. Cotransfection assays with a GlyRalpha2 minigene demonstrate that brPTB antagonizes the action of Nova to increase utilization of GlyRalpha2 E3A. brPTB binds to a 90-nt GlyRalpha2 RNA adjacent to the Nova binding site, but with an affinity that is more than 10-fold lower than Nova. When a putative binding site for brPTB on the GlyRalpha2 RNA is mutated, binding is abolished and the inhibitory effect on Nova-dependent exon selection disappears. These results suggest that brPTB is a tissue-restricted RNA binding protein that interacts with and inhibits the ability of Nova to activate exon selection in neurons.

MeSH Terms
Alternative Splicing Animals Antigens, Neoplasm/physiology Base Sequence Brain/metabolism Male Molecular Sequence Data Nerve Tissue Proteins/physiology Neuro-Oncological Ventral Antigen Polypyrimidine Tract-Binding Protein RNA-Binding Proteins/physiology Rats Rats, Sprague-Dawley Receptors, Glycine/genetics Ribonucleoproteins/physiology
Chemicals
Antigens, Neoplasm Nerve Tissue Proteins Neuro-Oncological Ventral Antigen RNA-Binding Proteins Receptors, Glycine Ribonucleoproteins Polypyrimidine Tract-Binding Protein
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Polydorides A D
Laboratory of Molecular Neuro-Oncology, The Rockefeller University, New York, NY 10021, USA.
Okano H J
Yang Y Y
Stefani G
Darnell R B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.110128397
Published
2000-06-06
Pages
6350-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC18606
Subset
IM
Grants
NINDS NIH HHS · R01 NS034389 · United States
NCI NIH HHS · T32 CA009673 · United States
NCI NIH HHS · CA 09673-18 · United States
NINDS NIH HHS · R01 NS34389 · United States
Databases
GENBANK
AF095718
External Links
PubMed source DOI: 10.1073/pnas.110128397 PMC Full Text View journal details
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