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Polypyrimidine tract-binding protein is involved in vivo in repression of a composite internal/3' -terminal exon of the Xenopus alpha-tropomyosin Pre-mRNA.
J Biol Chem. 2004 May 21; 279(21):22166-75.JB

Abstract

The Xenopus alpha(fast)-tropomyosin gene contains, at its 3' -end, a composite internal/3' -terminal exon (exon 9A9'), which is subjected to three different patterns of splicing according to the cell type. Exon 9A9' is included as a terminal exon in the myotome and as an internal exon in adult striated muscles, whereas it is skipped in nonmuscle cells. We have developed an in vivo model based on transient expression of minigenes encompassing the regulated exon 9A9' in Xenopus oocytes and embryos. We first show that the different alpha-tropomyosin minigenes recapitulate the splicing pattern of the endogenous gene and constitute valuable tools to seek regulatory sequences involved in exon 9A9' usage. A mutational analysis led to the identification of an intronic element that is involved in the repression of exon 9A9' in nonmuscle cells. This element harbors four polypyrimidine track-binding protein (PTB) binding sites that are essential for the repression of exon 9A9'. We show using UV cross-linking and immunoprecipitation experiments that Xenopus PTB (XPTB) interacts with these PTB binding sites. Finally, we show that depletion of endogenous XPTB in Xenopus embryos using a morpholinobased translational inhibition strategy resulted in exon 9A9' inclusion in embryonic epidermal cells. These results demonstrate that XPTB is required in vivo to repress the terminal exon 9A9' and suggest that PTB could be a major actor in the repression of regulated 3' -terminal exon.

Authors+Show Affiliations

UMR 6061 CNRS, Université de Rennes 1, Faculté de Médecine, 35043 Rennes cedex, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15010470

Citation

Hamon, Sandra, et al. "Polypyrimidine Tract-binding Protein Is Involved in Vivo in Repression of a Composite Internal/3' -terminal Exon of the Xenopus Alpha-tropomyosin Pre-mRNA." The Journal of Biological Chemistry, vol. 279, no. 21, 2004, pp. 22166-75.
Hamon S, Le Sommer C, Mereau A, et al. Polypyrimidine tract-binding protein is involved in vivo in repression of a composite internal/3' -terminal exon of the Xenopus alpha-tropomyosin Pre-mRNA. J Biol Chem. 2004;279(21):22166-75.
Hamon, S., Le Sommer, C., Mereau, A., Allo, M. R., & Hardy, S. (2004). Polypyrimidine tract-binding protein is involved in vivo in repression of a composite internal/3' -terminal exon of the Xenopus alpha-tropomyosin Pre-mRNA. The Journal of Biological Chemistry, 279(21), 22166-75.
Hamon S, et al. Polypyrimidine Tract-binding Protein Is Involved in Vivo in Repression of a Composite Internal/3' -terminal Exon of the Xenopus Alpha-tropomyosin Pre-mRNA. J Biol Chem. 2004 May 21;279(21):22166-75. PubMed PMID: 15010470.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Polypyrimidine tract-binding protein is involved in vivo in repression of a composite internal/3' -terminal exon of the Xenopus alpha-tropomyosin Pre-mRNA. AU - Hamon,Sandra, AU - Le Sommer,Caroline, AU - Mereau,Agnès, AU - Allo,Marie-Rose, AU - Hardy,Serge, Y1 - 2004/03/09/ PY - 2004/3/11/pubmed PY - 2004/7/1/medline PY - 2004/3/11/entrez SP - 22166 EP - 75 JF - The Journal of biological chemistry JO - J Biol Chem VL - 279 IS - 21 N2 - The Xenopus alpha(fast)-tropomyosin gene contains, at its 3' -end, a composite internal/3' -terminal exon (exon 9A9'), which is subjected to three different patterns of splicing according to the cell type. Exon 9A9' is included as a terminal exon in the myotome and as an internal exon in adult striated muscles, whereas it is skipped in nonmuscle cells. We have developed an in vivo model based on transient expression of minigenes encompassing the regulated exon 9A9' in Xenopus oocytes and embryos. We first show that the different alpha-tropomyosin minigenes recapitulate the splicing pattern of the endogenous gene and constitute valuable tools to seek regulatory sequences involved in exon 9A9' usage. A mutational analysis led to the identification of an intronic element that is involved in the repression of exon 9A9' in nonmuscle cells. This element harbors four polypyrimidine track-binding protein (PTB) binding sites that are essential for the repression of exon 9A9'. We show using UV cross-linking and immunoprecipitation experiments that Xenopus PTB (XPTB) interacts with these PTB binding sites. Finally, we show that depletion of endogenous XPTB in Xenopus embryos using a morpholinobased translational inhibition strategy resulted in exon 9A9' inclusion in embryonic epidermal cells. These results demonstrate that XPTB is required in vivo to repress the terminal exon 9A9' and suggest that PTB could be a major actor in the repression of regulated 3' -terminal exon. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/15010470/Polypyrimidine_tract_binding_protein_is_involved_in_vivo_in_repression_of_a_composite_internal/3'__terminal_exon_of_the_Xenopus_alpha_tropomyosin_Pre_mRNA_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=15010470 DB - PRIME DP - Unbound Medicine ER -