Binding of hnRNP H to an exonic splicing silencer is involved in the regulation of alternative splicing of the rat beta-tropomyosin gene

Chen, C. D., Kobayashi, R., Helfman, D. M. (March 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 and Development, 13 (5). pp. 593-606. ISSN 0890-9369 (Print)

Preview

PDF Kobayashi_Helfman_GenesDev1999.pdf Download (1MB) | Preview

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.

Item Type:	Paper
Uncontrolled Keywords:	Alternative Splicing Animals Base Sequence Exons Gene Expression Regulation Hela Cells Heterogeneous-Nuclear Ribonucleoprotein Group F-H Heterogeneous-Nuclear Ribonucleoproteins Humans Molecular Sequence Data Mutagenesis Rna Rats Recombinant Proteins/metabolism Research Support, U.S. Gov't, P.H.S. Ribonucleoproteins/ metabolism Tropomyosin/ genetics
Subjects:	bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Alternative Splicing bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > exons bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mRNA bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mutations > mutagenesis bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > nuclear ribonucleoprotein
CSHL Authors:	Helfman, David M. Kobayashi, Ryuji Chen, Degui
Communities:	CSHL labs > Helfman lab CSHL labs > Kobayashi lab
Depositing User:	Kathleen Darby
Date:	1 March 1999
Date Deposited:	30 Apr 2014 13:06
Last Modified:	30 Apr 2014 18:05
PMCID:	PMC316507
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/29798

Actions (login required)

Administrator's edit/view item