共 50 条
Identification of putative new splicing targets for ETR-3 using sequences identified by systematic evolution of ligands by exponential enrichment
被引:78
作者:

Faustino, NA
论文数: 0 引用数: 0
h-index: 0
机构: Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA

Cooper, TA
论文数: 0 引用数: 0
h-index: 0
机构: Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA
机构:
[1] Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA
[2] Baylor Coll Med, Dept Mol & Cell Biol, Houston, TX 77030 USA
[3] Univ Porto, Grad Program Basic & Appl Biol, ICBAS, Oporto, Portugal
关键词:
D O I:
10.1128/MCB.25.3.879-887.2005
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
ETR-3 (also know as BRUNOL3, NAPOR, and CUGBP2) is one of six members of the CELF (CUG-BP1- and ETR-3-like factor) family of splicing regulators. ETR-3 regulates splicing by direct binding to the pre-mRNA. We performed systematic evolution of ligands by exponential enrichment (SELEX) to identify the preferred binding sequence of ETR-3. After five rounds of SELEX, ETR-3 selected UG-rich sequences. in particular UG repeats and UGUU motifs. Either of these selected motifs was able to restore ETR-3 binding and responsiveness to a nonresponsive splicing reporter in vivo. Moreover, this effect was not specific to ETR-3 since minigenes containing either of the two motifs were responsive to two other CELF proteins (CUG-BP1 and CELF4), indicating that different members of the CELF family can mediate their effects via a common binding site. Using the SELEX-identified motifs to search the human genome. we identified several possible new ETR-3 targets. We created minigenes for two of these genes, the CFTR and MTMR1 genes. and confirmed that ETR-3 regulates their splicing patterns. For the CFTR minigene this regulation was demonstrated to be dependent on the presence of the putative binding site identified in our screen. These results validate this approach to search for new targets for RNA processing proteins.
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页码:879 / 887
页数:9
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