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Heterogeneous nuclear ribonucleoprotein (hnRNP) K is a component of an intronic splicing enhancer complex that activates the splicing of the alternative exon 6A from chicken beta-tropomyosin pre-mRNA.
J Biol Chem. 2002 May 10; 277(19):16614-23.JB

Abstract

Splicing of the chicken beta-tropomyosin exon 6A is stimulated, both in vivo and in vitro, by an intronic pyrimidine-rich element (S4) located 37 nucleotides downstream of exon 6A. Several pyrimidine-rich sequences are able to substitute for the natural S4 enhancer with various stimulatory effects. We show that the different enhancer sequences recruit U1 small nuclear ribonucleoprotein (SnRNP) to the exon 6A 5' splice site, with an efficiency that correlates with the splicing activation. By using RNA affinity and two-dimensional gel electrophoresis, we characterized several proteins that bind to the different enhancer sequences. Heterogeneous nuclear ribonucleoprotein (hnRNP) K and hnRNP I (polypyrimidine track-binding protein, PTB) exhibit a higher level of interaction with the strong enhancer sequences (S4) than with the weakest enhancers. Functional analysis shows that hnRNP K is a component of the enhancer complex that promotes exon 6A splicing through the wild-type S4 sequence. The addition of recombinant hnRNP K to nuclear extracts preincubated with poly(rC) RNA competitor completely restores splicing efficiency to the original level. hnRNP I (PTB) was also found associated with the strong enhancer sequences. Its function in the splicing of exon 6A is discussed.

Authors+Show Affiliations

Centre de Génétique Moléculaire, CNRS, F-91190 Gif-sur-Yvette, France.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11867641

Citation

Expert-Bezançon, Alain, et al. "Heterogeneous Nuclear Ribonucleoprotein (hnRNP) K Is a Component of an Intronic Splicing Enhancer Complex That Activates the Splicing of the Alternative Exon 6A From Chicken Beta-tropomyosin Pre-mRNA." The Journal of Biological Chemistry, vol. 277, no. 19, 2002, pp. 16614-23.
Expert-Bezançon A, Le Caer JP, Marie J. Heterogeneous nuclear ribonucleoprotein (hnRNP) K is a component of an intronic splicing enhancer complex that activates the splicing of the alternative exon 6A from chicken beta-tropomyosin pre-mRNA. J Biol Chem. 2002;277(19):16614-23.
Expert-Bezançon, A., Le Caer, J. P., & Marie, J. (2002). Heterogeneous nuclear ribonucleoprotein (hnRNP) K is a component of an intronic splicing enhancer complex that activates the splicing of the alternative exon 6A from chicken beta-tropomyosin pre-mRNA. The Journal of Biological Chemistry, 277(19), 16614-23.
Expert-Bezançon A, Le Caer JP, Marie J. Heterogeneous Nuclear Ribonucleoprotein (hnRNP) K Is a Component of an Intronic Splicing Enhancer Complex That Activates the Splicing of the Alternative Exon 6A From Chicken Beta-tropomyosin Pre-mRNA. J Biol Chem. 2002 May 10;277(19):16614-23. PubMed PMID: 11867641.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Heterogeneous nuclear ribonucleoprotein (hnRNP) K is a component of an intronic splicing enhancer complex that activates the splicing of the alternative exon 6A from chicken beta-tropomyosin pre-mRNA. AU - Expert-Bezançon,Alain, AU - Le Caer,Jean Pierre, AU - Marie,Joëlle, Y1 - 2002/02/26/ PY - 2002/2/28/pubmed PY - 2002/6/14/medline PY - 2002/2/28/entrez SP - 16614 EP - 23 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 277 IS - 19 N2 - Splicing of the chicken beta-tropomyosin exon 6A is stimulated, both in vivo and in vitro, by an intronic pyrimidine-rich element (S4) located 37 nucleotides downstream of exon 6A. Several pyrimidine-rich sequences are able to substitute for the natural S4 enhancer with various stimulatory effects. We show that the different enhancer sequences recruit U1 small nuclear ribonucleoprotein (SnRNP) to the exon 6A 5' splice site, with an efficiency that correlates with the splicing activation. By using RNA affinity and two-dimensional gel electrophoresis, we characterized several proteins that bind to the different enhancer sequences. Heterogeneous nuclear ribonucleoprotein (hnRNP) K and hnRNP I (polypyrimidine track-binding protein, PTB) exhibit a higher level of interaction with the strong enhancer sequences (S4) than with the weakest enhancers. Functional analysis shows that hnRNP K is a component of the enhancer complex that promotes exon 6A splicing through the wild-type S4 sequence. The addition of recombinant hnRNP K to nuclear extracts preincubated with poly(rC) RNA competitor completely restores splicing efficiency to the original level. hnRNP I (PTB) was also found associated with the strong enhancer sequences. Its function in the splicing of exon 6A is discussed. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/11867641/Heterogeneous_nuclear_ribonucleoprotein__hnRNP__K_is_a_component_of_an_intronic_splicing_enhancer_complex_that_activates_the_splicing_of_the_alternative_exon_6A_from_chicken_beta_tropomyosin_pre_mRNA_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=11867641 DB - PRIME DP - Unbound Medicine ER -