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Binding of hnRNP H to an exonic splicing silencer is involved in the regulation of alternative splicing of the rat beta-tropomyosin gene.
Genes Dev. 1999 Mar 01; 13(5):593-606.GD

Abstract

In the rat beta-tropomyosin (beta-TM) gene, exons 6 and 7 are spliced alternatively in a mutually exclusive manner. Exon 6 is included in mRNA encoding nonmuscle TM-1, whereas exon 7 is used in mRNA encoding skeletal muscle beta-TM. Previously, we demonstrated that a six nucleotide mutation at the 5' end of exon 7, designated as ex-1, activated exon 7 splicing in nonmuscle cells. In this study, we show that the activating effect of this mutation is not the result of creating an exonic splicing enhancer (ESE) or disrupting a putative secondary structure. The sequence in exon 7 acts as a bona fide exonic splicing silencer (ESS), which is bound specifically by a trans-acting factor. Isolation and peptide sequencing reveal that this factor is hnRNP H, a member of the heterogeneous nuclear ribonucleoprotein (hnRNP) family. Binding of hnRNP H correlates with the ESS activity. Furthermore, addition of antibodies that specifically recognizes hnRNP H to the splicing reactions or partial depletion of hnRNP H from nuclear extract activates exon 7 splicing in vitro and this effect can be reversed by addition of purified recombinant hnRNP H. These results indicate that hnRNP H participates in exclusion of exon 7 in nonmuscle cells. The involvement of hnRNP H in the activity of an ESS may represent a prototype for the regulation of tissue- and developmental-specific alternative splicing.

Authors+Show Affiliations

Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

10072387

Citation

Chen, C D., et al. "Binding of hnRNP H to an Exonic Splicing Silencer Is Involved in the Regulation of Alternative Splicing of the Rat Beta-tropomyosin Gene." Genes & Development, vol. 13, no. 5, 1999, pp. 593-606.
Chen CD, Kobayashi R, Helfman DM. Binding of hnRNP H to an exonic splicing silencer is involved in the regulation of alternative splicing of the rat beta-tropomyosin gene. Genes Dev. 1999;13(5):593-606.
Chen, C. D., Kobayashi, R., & Helfman, D. M. (1999). Binding of hnRNP H to an exonic splicing silencer is involved in the regulation of alternative splicing of the rat beta-tropomyosin gene. Genes & Development, 13(5), 593-606.
Chen CD, Kobayashi R, Helfman DM. Binding of hnRNP H to an Exonic Splicing Silencer Is Involved in the Regulation of Alternative Splicing of the Rat Beta-tropomyosin Gene. Genes Dev. 1999 Mar 1;13(5):593-606. PubMed PMID: 10072387.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Binding of hnRNP H to an exonic splicing silencer is involved in the regulation of alternative splicing of the rat beta-tropomyosin gene. AU - Chen,C D, AU - Kobayashi,R, AU - Helfman,D M, PY - 1999/3/11/pubmed PY - 1999/3/11/medline PY - 1999/3/11/entrez SP - 593 EP - 606 JF - Genes & development JO - Genes Dev VL - 13 IS - 5 N2 - In the rat beta-tropomyosin (beta-TM) gene, exons 6 and 7 are spliced alternatively in a mutually exclusive manner. Exon 6 is included in mRNA encoding nonmuscle TM-1, whereas exon 7 is used in mRNA encoding skeletal muscle beta-TM. Previously, we demonstrated that a six nucleotide mutation at the 5' end of exon 7, designated as ex-1, activated exon 7 splicing in nonmuscle cells. In this study, we show that the activating effect of this mutation is not the result of creating an exonic splicing enhancer (ESE) or disrupting a putative secondary structure. The sequence in exon 7 acts as a bona fide exonic splicing silencer (ESS), which is bound specifically by a trans-acting factor. Isolation and peptide sequencing reveal that this factor is hnRNP H, a member of the heterogeneous nuclear ribonucleoprotein (hnRNP) family. Binding of hnRNP H correlates with the ESS activity. Furthermore, addition of antibodies that specifically recognizes hnRNP H to the splicing reactions or partial depletion of hnRNP H from nuclear extract activates exon 7 splicing in vitro and this effect can be reversed by addition of purified recombinant hnRNP H. These results indicate that hnRNP H participates in exclusion of exon 7 in nonmuscle cells. The involvement of hnRNP H in the activity of an ESS may represent a prototype for the regulation of tissue- and developmental-specific alternative splicing. SN - 0890-9369 UR - https://www.unboundmedicine.com/medline/citation/10072387/Binding_of_hnRNP_H_to_an_exonic_splicing_silencer_is_involved_in_the_regulation_of_alternative_splicing_of_the_rat_beta_tropomyosin_gene_ L2 - http://www.genesdev.org/cgi/pmidlookup?view=long&pmid=10072387 DB - PRIME DP - Unbound Medicine ER -