論文 - 詳細
| RRC ID | 53269 |
|---|---|
| 著者 | Itani OA, Flibotte S, Dumas KJ, Guo C, Blumenthal T, Hu PJ. |
| タイトル | N-Ethyl-N-Nitrosourea (ENU) Mutagenesis Reveals an Intronic Residue Critical for Caenorhabditis elegans 3' Splice Site Function in Vivo. |
| ジャーナル | G3 (Bethesda) |
| Abstract |
Metazoan introns contain a polypyrimidine tract immediately upstream of the AG dinucleotide that defines the 3' splice site. In the nematode Caenorhabditis elegans, 3' splice sites are characterized by a highly conserved UUUUCAG/R octamer motif. While the conservation of pyrimidines in this motif is strongly suggestive of their importance in pre-mRNA splicing, in vivo evidence in support of this is lacking. In an N-ethyl-N-nitrosourea (ENU) mutagenesis screen in Caenorhabditis elegans, we have isolated a strain containing a point mutation in the octamer motif of a 3' splice site in the daf-12 gene. This mutation, a single base T-to-G transversion at the -5 position relative to the splice site, causes a strong daf-12 loss-of-function phenotype by abrogating splicing. The resulting transcript is predicted to encode a truncated DAF-12 protein generated by translation into the retained intron, which contains an in-frame stop codon. Other than the perfectly conserved AG dinucleotide at the site of splicing, G at the -5 position of the octamer motif is the most uncommon base in C. elegans 3' splice sites, occurring at closely paired sites where the better match to the splicing consensus is a few bases downstream. Our results highlight both the biological importance of the highly conserved -5 uridine residue in the C. elegans 3' splice site octamer motif as well as the utility of using ENU as a mutagen to study the function of polypyrimidine tracts and other AU- or AT-rich motifs in vivo. |
| 巻・号 | 6(6) |
| ページ | 1751-6 |
| 公開日 | 2016-6-1 |
| DOI | 10.1534/g3.116.028662 |
| PII | g3.116.028662 |
| PMID | 27172199 |
| PMC | PMC4889670 |
| MeSH | Animals Base Sequence Caenorhabditis elegans / drug effects* Caenorhabditis elegans / genetics* Caenorhabditis elegans / metabolism Chromosome Mapping Ethylnitrosourea / toxicity* Introns* Mutagenesis / drug effects* Mutation Nucleotide Motifs Phenotype Polymorphism, Single Nucleotide Protein Binding RNA Splice Sites* RNA Splicing Factors / metabolism Sequence Deletion |
| IF | 2.781 |
| 引用数 | 1 |
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オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 6 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 線虫 | tm3188 |