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Title: Crystallization and preliminary X-ray analysis of a U2AF{sup 65} variant in complex with a polypyrimidine-tract analogue by use of protein engineering

Abstract

A complex of the essential splicing factor U2AF{sup 65} and a deoxyuridine oligonucleotide has been crystallized by modification of an interdomain linker. The large subunit of the essential pre-mRNA splicing factor U2 auxiliary factor (U2AF{sup 65}) binds the polypyrimidine tract near the 3′ splice site of pre-mRNA introns and directs the association of the U2 small nuclear ribonucleoprotein particle (U2 snRNP) of the spliceosome with the pre-mRNA. Protein engineering, in which the flexible linker region connecting tandem RNA-recognition motifs (RRMs) within the U2AF{sup 65} RNA-binding domain was partially deleted, allowed successful crystallization of the protein–nucleic acid complex. Cocrystals of a U2AF{sup 65} variant with a deoxyuridine dodecamer diffract X-rays to 2.9 Å resolution and contain one complex per asymmetric unit.

Authors:
;
Publication Date:
OSTI Identifier:
22356318
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section F
Additional Journal Information:
Journal Volume: 62; Journal Issue: Pt 5; Other Information: PMCID: PMC2219975; PMID: 16682775; PUBLISHER-ID: ll5055; OAI: oai:pubmedcentral.nih.gov:2219975; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 1744-3091
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; FASTENING; MODIFICATIONS; PROTEINS; RESOLUTION

Citation Formats

Sickmier, E. Allen, Frato, Katherine E., and Kielkopf, Clara L., E-mail: ckielkop@jhsph.edu. Crystallization and preliminary X-ray analysis of a U2AF{sup 65} variant in complex with a polypyrimidine-tract analogue by use of protein engineering. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106012504.
Sickmier, E. Allen, Frato, Katherine E., & Kielkopf, Clara L., E-mail: ckielkop@jhsph.edu. Crystallization and preliminary X-ray analysis of a U2AF{sup 65} variant in complex with a polypyrimidine-tract analogue by use of protein engineering. United Kingdom. https://doi.org/10.1107/S1744309106012504
Sickmier, E. Allen, Frato, Katherine E., and Kielkopf, Clara L., E-mail: ckielkop@jhsph.edu. 2006. "Crystallization and preliminary X-ray analysis of a U2AF{sup 65} variant in complex with a polypyrimidine-tract analogue by use of protein engineering". United Kingdom. https://doi.org/10.1107/S1744309106012504.
@article{osti_22356318,
title = {Crystallization and preliminary X-ray analysis of a U2AF{sup 65} variant in complex with a polypyrimidine-tract analogue by use of protein engineering},
author = {Sickmier, E. Allen and Frato, Katherine E. and Kielkopf, Clara L., E-mail: ckielkop@jhsph.edu},
abstractNote = {A complex of the essential splicing factor U2AF{sup 65} and a deoxyuridine oligonucleotide has been crystallized by modification of an interdomain linker. The large subunit of the essential pre-mRNA splicing factor U2 auxiliary factor (U2AF{sup 65}) binds the polypyrimidine tract near the 3′ splice site of pre-mRNA introns and directs the association of the U2 small nuclear ribonucleoprotein particle (U2 snRNP) of the spliceosome with the pre-mRNA. Protein engineering, in which the flexible linker region connecting tandem RNA-recognition motifs (RRMs) within the U2AF{sup 65} RNA-binding domain was partially deleted, allowed successful crystallization of the protein–nucleic acid complex. Cocrystals of a U2AF{sup 65} variant with a deoxyuridine dodecamer diffract X-rays to 2.9 Å resolution and contain one complex per asymmetric unit.},
doi = {10.1107/S1744309106012504},
url = {https://www.osti.gov/biblio/22356318}, journal = {Acta Crystallographica. Section F},
issn = {1744-3091},
number = Pt 5,
volume = 62,
place = {United Kingdom},
year = {Mon May 01 00:00:00 EDT 2006},
month = {Mon May 01 00:00:00 EDT 2006}
}