DC Field | Value | Language |
---|---|---|
dc.contributor.author | Selvakumar, Meenakshi | ko |
dc.contributor.author | Helfman, David M | ko |
dc.date.accessioned | 2013-03-02T13:19:13Z | - |
dc.date.available | 2013-03-02T13:19:13Z | - |
dc.date.created | 2012-02-06 | - |
dc.date.created | 2012-02-06 | - |
dc.date.issued | 1999-03 | - |
dc.identifier.citation | RNA-A PUBLICATION OF THE RNA SOCIETY, v.5, no.3, pp.378 - 394 | - |
dc.identifier.issn | 1355-8382 | - |
dc.identifier.uri | http://hdl.handle.net/10203/73710 | - |
dc.description.abstract | The rat beta-tropomyosin gene encodes two tissue-specific isoforms that contain the internal, mutually exclusive exons 6 (nonmuscle/smooth muscle) and 7 (skeletal muscle). We previously demonstrated that the 3' splice site of exon 6 can be activated by introducing a 9-nt polyuridine tract at its 3' splice site, or by strengthening the 5' splice site to a U1 consensus binding site, or by joining exon 6 to the downstream common exon 8. Examination of sequences within exons 6 and 8 revealed the presence of two purine-rich motifs in exon 6 and three purine-rich motifs in exon 8 that could potentially represent exonic splicing enhancers (ESEs). In this report we carried out substitution mutagenesis of these elements and show that some of them play a critical role in the splice site usage of exon 6 in vitro and in vivo. Using UV crosslinking, we have identified SF2/ASF as one of the cellular factors that binds to these motifs. Furthermore, we show that substrates that have mutated ESEs are blocked prior to A-complex formation, supporting a role for SF2/ASF binding to the ESEs during the commitment step in splicing. Using pre-mRNA substrates containing exons 5 through 8, we show that the ESEs within exon 6 also play a role in cooperation between the 3' and 5' splice sites flanking this exon. The splicing of exon 6 to 8 (i.e., 5' splice site usage of exon 6) was enhanced with pre-mRNAs containing either the polyuridine tract in the 3' splice site or consensus sequence in the 5' splice site around exon 6. We show that the ESEs in exon 6 are required for this effect. However, the ESEs are not required when both the polyuridine and consensus splice site sequences around exon 6 were present in the same pre-mRNA. These results support and extend the exon-definition hypothesis and demonstrate that sequences at the 3' splice site can facilitate use of a downstream 5' splice site. In addition, the data support the hypothesis that ESEs can compensate for weak splice sites, such as those found in alternatively spliced exons, thereby providing a target for regulation. | - |
dc.language | English | - |
dc.publisher | CAMBRIDGE UNIV PRESS | - |
dc.subject | T ALTERNATIVE EXON | - |
dc.subject | MESSENGER-RNA | - |
dc.subject | SR-PROTEINS | - |
dc.subject | PREMESSENGER RNA | - |
dc.subject | NONMUSCLE CELLS | - |
dc.subject | FACTORS ASF/SF2 | - |
dc.subject | CIS-ELEMENTS | - |
dc.subject | SELECTION | - |
dc.subject | DISTINCT | - |
dc.subject | BINDING | - |
dc.title | Exonic splicing enhancers contribute to the use of both 3 and 5 splice site usage of rat beta-tropomyosin pre-mRNA | - |
dc.type | Article | - |
dc.identifier.wosid | 000079007200005 | - |
dc.identifier.scopusid | 2-s2.0-0033018827 | - |
dc.type.rims | ART | - |
dc.citation.volume | 5 | - |
dc.citation.issue | 3 | - |
dc.citation.beginningpage | 378 | - |
dc.citation.endingpage | 394 | - |
dc.citation.publicationname | RNA-A PUBLICATION OF THE RNA SOCIETY | - |
dc.identifier.doi | 10.1017/S1355838299981050 | - |
dc.contributor.localauthor | Helfman, David M | - |
dc.contributor.nonIdAuthor | Selvakumar, Meenakshi | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | E complex | - |
dc.subject.keywordAuthor | exon definition | - |
dc.subject.keywordAuthor | purine-rich motifs | - |
dc.subject.keywordAuthor | SF2/ASF | - |
dc.subject.keywordPlus | T ALTERNATIVE EXON | - |
dc.subject.keywordPlus | MESSENGER-RNA | - |
dc.subject.keywordPlus | SR-PROTEINS | - |
dc.subject.keywordPlus | PREMESSENGER RNA | - |
dc.subject.keywordPlus | NONMUSCLE CELLS | - |
dc.subject.keywordPlus | FACTORS ASF/SF2 | - |
dc.subject.keywordPlus | CIS-ELEMENTS | - |
dc.subject.keywordPlus | SELECTION | - |
dc.subject.keywordPlus | DISTINCT | - |
dc.subject.keywordPlus | BINDING | - |
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