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Related Experiment Video

Updated: Jul 12, 2026

Mapping the Binding Site of an Aptamer on ATP Using MicroScale Thermophoresis

Mapping the Binding Site of an Aptamer on ATP Using MicroScale Thermophoresis

Published on: January 8, 2017

Distinct binding specificities and functions of higher eukaryotic polypyrimidine tract-binding proteins

R Singh¹, J Valcárcel, M R Green

¹Howard Hughes Medical Institute, Program in Molecular Medicine, University of Massachusetts Medical Center, Worcester 01605, USA.

Science (New York, N.Y.)

|May 26, 1995

Summary

Polypyrimidine tract-binding protein (PTB) and other splicing factors recognize distinct sequences near splice sites. PTB selectively represses alternative splicing by binding to specific sites, influencing gene expression.

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Last Updated: Jul 12, 2026

Mapping the Binding Site of an Aptamer on ATP Using MicroScale Thermophoresis

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Published on: January 8, 2017

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

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DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling

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Published on: October 8, 2019

Area of Science:

Molecular Biology
Genetics
RNA Splicing

Background:

The polypyrimidine tract (Py-tract) near the 3' splice site is crucial for pre-mRNA splicing in eukaryotes.
Several proteins, including U2AF65, Sex-lethal (Sxl), and polypyrimidine tract-binding protein (PTB), interact with the Py-tract.
The precise function and sequence preferences of PTB in splicing regulation were previously unknown.

Purpose of the Study:

To investigate the distinct sequence preferences of splicing factors U2AF65, Sxl, and PTB.
To elucidate the role of PTB in alternative splicing regulation.

Main Methods:

Iterative in vitro genetic selection was employed to determine protein-specific sequence preferences.
Analysis of natural metazoan Py-tracts and alternatively spliced pre-mRNAs.

Main Results:

U2AF65 binds to uridine-rich sequences similar to natural metazoan Py-tracts.
Sxl recognizes a highly specific consensus sequence, explaining its regulatory roles.
PTB binds to a distinct consensus sequence found in negatively regulated alternatively spliced pre-mRNAs.
PTB was shown to selectively repress 3' splice sites containing its binding site.

Conclusions:

Splicing factors exhibit distinct sequence specificities for Py-tract recognition.
PTB plays a significant role in alternative splicing by repressing specific 3' splice sites.
Understanding these interactions provides insights into the regulation of gene expression through alternative splicing.