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Strong RNA splicing enhancers identified by a modified method of cycled selection interact with SR protein.
J Biol Chem. 2001 Sep 07; 276(36):33833-9.JB

Abstract

A modified method of cycled selection was used to characterize splicing enhancers for exon inclusion from a pool of beta-globin-based three exon/two intron pre-mRNAs with a variable number of random nucleotides incorporated in the internal exon. The pre-mRNAs generated by this method contained random sequences ranging from 0 to 18 nucleotides in length. This method was used to isolate particular splicing enhancer motifs from a previously enriched pool of extremely diverse enhancers. After four cycles of selection for mRNA containing the internal exon, a distinct enhancer motif (GACGAC...CAGCAG) was highly enriched. This motif served as strong splicing enhancers in a heterogeneous exon. We have shown here that the selected enhancer motif promotes exon inclusion through specific interaction with SRp30. We have also shown that although present in many of our selected splicing enhancers conforming to this motif, a typical purine-rich enhancer sequence is dispensable for either enhancer activity or binding with SRp30.

Links

Publisher Full Text
linkinghub.elsevier.com
jbc.org

Authors+Show Affiliations

Tian H
Department of Genetics, Center for Genetic and Cellular Therapies, Duke University, Durham, North Carolina 27710, USA. h.tian@cgct.duke.edu
Kole R
No affiliation info available

MeSH

Amino Acid MotifsBase SequenceChromatography, AffinityCross-Linking ReagentsEnhancer Elements, GeneticEscherichia coliExonsHeLa CellsHumansIntronsModels, GeneticMolecular Sequence DataNuclear ProteinsPhosphoproteinsPlasmidsProtein BindingRNA SplicingRNA, MessengerRNA-Binding ProteinsSerine-Arginine Splicing FactorsTranscription, GeneticUltraviolet Rays

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11454855

Citation

Tian, H, and R Kole. "Strong RNA Splicing Enhancers Identified By a Modified Method of Cycled Selection Interact With SR Protein." The Journal of Biological Chemistry, vol. 276, no. 36, 2001, pp. 33833-9.
Tian H, Kole R. Strong RNA splicing enhancers identified by a modified method of cycled selection interact with SR protein. J Biol Chem. 2001;276(36):33833-9.
Tian, H., & Kole, R. (2001). Strong RNA splicing enhancers identified by a modified method of cycled selection interact with SR protein. The Journal of Biological Chemistry, 276(36), 33833-9.
Tian H, Kole R. Strong RNA Splicing Enhancers Identified By a Modified Method of Cycled Selection Interact With SR Protein. J Biol Chem. 2001 Sep 7;276(36):33833-9. PubMed PMID: 11454855.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Strong RNA splicing enhancers identified by a modified method of cycled selection interact with SR protein. AU - Tian,H, AU - Kole,R, Y1 - 2001/07/13/ PY - 2001/7/17/pubmed PY - 2001/10/12/medline PY - 2001/7/17/entrez SP - 33833 EP - 9 JF - The Journal of biological chemistry JO - J Biol Chem VL - 276 IS - 36 N2 - A modified method of cycled selection was used to characterize splicing enhancers for exon inclusion from a pool of beta-globin-based three exon/two intron pre-mRNAs with a variable number of random nucleotides incorporated in the internal exon. The pre-mRNAs generated by this method contained random sequences ranging from 0 to 18 nucleotides in length. This method was used to isolate particular splicing enhancer motifs from a previously enriched pool of extremely diverse enhancers. After four cycles of selection for mRNA containing the internal exon, a distinct enhancer motif (GACGAC...CAGCAG) was highly enriched. This motif served as strong splicing enhancers in a heterogeneous exon. We have shown here that the selected enhancer motif promotes exon inclusion through specific interaction with SRp30. We have also shown that although present in many of our selected splicing enhancers conforming to this motif, a typical purine-rich enhancer sequence is dispensable for either enhancer activity or binding with SRp30. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/11454855/Strong_RNA_splicing_enhancers_identified_by_a_modified_method_of_cycled_selection_interact_with_SR_protein_ L2 - https://linkinghub.elsevier.com/retrieve/pii/M102957200 DB - PRIME DP - Unbound Medicine ER -