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Specific SR protein-dependent splicing substrates identified through genomic SELEX.

ABSTRACT: The Drosophila pre-mRNA splicing factor B52 (SRp55) is essential for fly development, but splicing of RNAs of specific genes tested previously is normal in B52-null animals, presumably due to partial functional redundancy with other SR proteins. To identify B52-dependent splicing substrates in vivo, we selected genomic sequence fragments whose transcripts bind B52. Almost all of the corresponding genes having a known function encode either transcription factors or components of signal transduction pathways, with the B52- binding fragments located to not only exonic but also intronic regions. Some pre-mRNAs from these genes showed splicing defects in the B52-null mutant. These results indicate that B52 has unique functions in the removal of some introns during development, and plays a critical role in cellular regulatory networks.

SUBMITTER: Kim S

PROVIDER: S-EPMC152802 | biostudies-literature | 2003 Apr

REPOSITORIES: biostudies-literature

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Specific SR protein-dependent splicing substrates identified through genomic SELEX.

Kim Soyoun S Shi Hua H Lee Dong-Ki DK Lis John T JT

Nucleic acids research 20030401 7

The Drosophila pre-mRNA splicing factor B52 (SRp55) is essential for fly development, but splicing of RNAs of specific genes tested previously is normal in B52-null animals, presumably due to partial functional redundancy with other SR proteins. To identify B52-dependent splicing substrates in vivo, we selected genomic sequence fragments whose transcripts bind B52. Almost all of the corresponding genes having a known function encode either transcription factors or components of signal transducti ...[more]

PMID: 12655012

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Project description:BackgroundSeveral recent studies indicate that alternative splicing in Arabidopsis and other plants is a common mechanism for post-transcriptional modulation of gene expression. However, few analyses have been done so far to elucidate the functional relevance of alternative splicing in higher plants. Representing a frequent and universal subtle alternative splicing event among eukaryotes, alternative splicing at NAGNAG acceptors contributes to transcriptome diversity and therefore, proteome plasticity. Alternatively spliced NAGNAG acceptors are overrepresented in genes coding for proteins with RNA-recognition motifs (RRMs). As SR proteins, a family of RRM-containing important splicing factors, are known to be extensively alternatively spliced in Arabidopsis, we analyzed alternative splicing at NAGNAG acceptors in SR and SR-related genes.ResultsIn a comprehensive analysis of the Arabidopsis thaliana genome, we identified 6,772 introns that exhibit a NAGNAG acceptor motif. Alternative splicing at these acceptors was assessed using available EST data, complemented by a sequence-based prediction method. Of the 36 identified introns within 30 SR and SR-related protein-coding genes that have a NAGNAG acceptor, we selected 15 candidates for an experimental analysis of alternative splicing under several conditions. We provide experimental evidence for 8 of these candidates being alternatively spliced. Quantifying the ratio of NAGNAG-derived splice variants under several conditions, we found organ-specific splicing ratios in adult plants and changes in seedlings of different ages. Splicing ratio changes were observed in response to heat shock and most strikingly, cold shock. Interestingly, the patterns of differential splicing ratios are similar for all analyzed genes.ConclusionNAGNAG acceptors frequently occur in the Arabidopsis genome and are particularly prevalent in SR and SR-related protein-coding genes. A lack of extensive EST coverage can be compensated by using the proposed sequence-based method to predict alternative splicing at these acceptors. Our findings indicate that the differential effects on NAGNAG alternative splicing in SR and SR-related genes are organ- and condition-specific rather than gene-specific.

Dataset Information

Specific SR protein-dependent splicing substrates identified through genomic SELEX.

Publications

Specific SR protein-dependent splicing substrates identified through genomic SELEX.

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