A strong exonic splicing enhancer in dystrophin exon 19 achieve proper splicing without an upstream polypyrimidine tract

被引：13

作者：

Habara, Yasuaki ^{[1
]}

Doshita, Mari ^{[2
]}

Hirozawa, Sadako ^{[2
]}

Yokono, Yuka ^{[2
]}

Yagi, Mariko ^{[1
]}

Takeshima, Yasuhiro ^{[1
]}

Matsuo, Masafumi ^{[1
]}

机构：

[1] Kobe Univ, Grad Sch Med, Dept Pediat, Kobe, Hyogo 6500017, Japan

[2] Kobe Pharmaceut Univ, Kobe, Hyogo 6588558, Japan

来源：

JOURNAL OF BIOCHEMISTRY | 2008年 / 143卷 / 03期

关键词：

acceptor site sequences; dystrophin; exonic splicing enhancer; polypyrimidine tract; splicing;

D O I：

10.1093/jb/mvm227

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

Proper splicing is known to proceed under the control of conserved cis-elements located at exon-intron boundaries. Recently, it was shown that additional elements, such as exonic splicing enhancers (ESEs), are essential for the proper splicing of certain exons, in addition to the splice donor and acceptor site sequences; however, the relationship between these cis-elements is still unclear. In this report, we utilize dystrophin exon 19 to analyse the relationship between the ESE and its upstream acceptor site sequences. Dystrophin exon 19, which maintains adequate splicing donor and acceptor consensus sequences, encodes exonic splicing enhancer (dys-ESE19) sequences. Splice pattern analysis, using a minigene reporter expressed in HeLa cells, showed that either a strong polypyrimidine tract (PPT) or a fully active dys-ESE19 is sufficient for proper splicing. Each of these two cis-elements has enough activity for proper exon 19 splicing suggesting that the PPT, which is believed to be an essential cis-element for splicing, is dispensable when the downstream exon contains a strong ESE. This compensation was only seen in living cells but not in 'in vitro splicing'. This suggests the possibility that the previous splicing experiments using an in vitro splicing system could underestimate the activity of ESEs.

引用

页码：303 / 310

页数：8

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