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Regulated splicing of an alternative exon of beta-tropomyosin pre-mRNAs in myogenic cells depends on the strength of pyrimidine-rich intronic enhancer elements.
DNA Cell Biol. 1999 Sep; 18(9):671-83.DC

Abstract

Alternative splicing of chicken beta-tropomyosin (beta-TM) pre-mRNAs ensures that in nonmuscle cells, only exon 6A is expressed, whereas in skeletal muscle, exon 6B is utilized preferentially. We have previously shown that efficient splicing of the nonmuscle exon 6A requires two pyrimidine-rich splicing enhancers (S4 and I5Y) that are present in the introns flanking exon 6A. Here, we examined the function of the S4 and I5Y elements by replacing them within beta-TM minigenes by other pyrimidine- and purine-rich sequence elements and analyzing splicing in transfected quail nonmuscle and muscle cells. Several features of these splicing regulatory elements were revealed by this study. First, a wide variety of pyrimidine-rich sequences can replace the intronic S4 splicing enhancer, indicating that pyrimidine composition, rather than sequence specificity, determines activity for this element. Second, one type of purine-rich sequence (GARn), normally found within exons, can also replace the S4 splicing enhancer. Third, the diverse elements tested exhibit differential activation of the splice sites flanking exon 6A and different positional constraints. Fourth, the strength of the S4 splicing enhancer is appropriately set to obtain proper regulation of the transition from exon 6A splicing in myoblasts to exon 6B splicing in myotubes, but this splicing regulatory element is not the target for cell-type-specific splicing factors.

Authors+Show Affiliations

INSERM U523, Groupe Hospitalier Pitié-Salpétrière, Paris, France.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

10492398

Citation

Pret, A M., et al. "Regulated Splicing of an Alternative Exon of Beta-tropomyosin pre-mRNAs in Myogenic Cells Depends On the Strength of Pyrimidine-rich Intronic Enhancer Elements." DNA and Cell Biology, vol. 18, no. 9, 1999, pp. 671-83.
Pret AM, Balvay L, Fiszman MY. Regulated splicing of an alternative exon of beta-tropomyosin pre-mRNAs in myogenic cells depends on the strength of pyrimidine-rich intronic enhancer elements. DNA Cell Biol. 1999;18(9):671-83.
Pret, A. M., Balvay, L., & Fiszman, M. Y. (1999). Regulated splicing of an alternative exon of beta-tropomyosin pre-mRNAs in myogenic cells depends on the strength of pyrimidine-rich intronic enhancer elements. DNA and Cell Biology, 18(9), 671-83.
Pret AM, Balvay L, Fiszman MY. Regulated Splicing of an Alternative Exon of Beta-tropomyosin pre-mRNAs in Myogenic Cells Depends On the Strength of Pyrimidine-rich Intronic Enhancer Elements. DNA Cell Biol. 1999;18(9):671-83. PubMed PMID: 10492398.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Regulated splicing of an alternative exon of beta-tropomyosin pre-mRNAs in myogenic cells depends on the strength of pyrimidine-rich intronic enhancer elements. AU - Pret,A M, AU - Balvay,L, AU - Fiszman,M Y, PY - 1999/9/24/pubmed PY - 1999/9/24/medline PY - 1999/9/24/entrez SP - 671 EP - 83 JF - DNA and cell biology JO - DNA Cell Biol VL - 18 IS - 9 N2 - Alternative splicing of chicken beta-tropomyosin (beta-TM) pre-mRNAs ensures that in nonmuscle cells, only exon 6A is expressed, whereas in skeletal muscle, exon 6B is utilized preferentially. We have previously shown that efficient splicing of the nonmuscle exon 6A requires two pyrimidine-rich splicing enhancers (S4 and I5Y) that are present in the introns flanking exon 6A. Here, we examined the function of the S4 and I5Y elements by replacing them within beta-TM minigenes by other pyrimidine- and purine-rich sequence elements and analyzing splicing in transfected quail nonmuscle and muscle cells. Several features of these splicing regulatory elements were revealed by this study. First, a wide variety of pyrimidine-rich sequences can replace the intronic S4 splicing enhancer, indicating that pyrimidine composition, rather than sequence specificity, determines activity for this element. Second, one type of purine-rich sequence (GARn), normally found within exons, can also replace the S4 splicing enhancer. Third, the diverse elements tested exhibit differential activation of the splice sites flanking exon 6A and different positional constraints. Fourth, the strength of the S4 splicing enhancer is appropriately set to obtain proper regulation of the transition from exon 6A splicing in myoblasts to exon 6B splicing in myotubes, but this splicing regulatory element is not the target for cell-type-specific splicing factors. SN - 1044-5498 UR - https://www.unboundmedicine.com/medline/citation/10492398/Regulated_splicing_of_an_alternative_exon_of_beta_tropomyosin_pre_mRNAs_in_myogenic_cells_depends_on_the_strength_of_pyrimidine_rich_intronic_enhancer_elements_ L2 - https://www.liebertpub.com/doi/10.1089/104454999314953?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -