Unknown,Transcriptomics,Genomics,Proteomics

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Mediator Complex Regulates Alternative mRNA Processing via the Med23 Subunit


ABSTRACT: Mediator complex is an integrative hub for transcriptional regulation. Here we show that Mediator regulates alternative mRNA processing via its Med23 subunit. Combining tandem affinity purification and mass spectrometry, we identified a number of mRNA processing factors that bind to a soluble recombinant Mediator subunit MED23 but not to several other Mediator components. One of these factors, hnRNP L, specifically interacts with MED23 in vitro and in vivo. Consistently, Mediator partially colocalizes with hnRNP L and the splicing machinery in the cell. Functionally Med23 regulates a subset of hnRNP L-targeted alternative splicing (AS) and alternative cleavage and polyadenylation (APA) events as shown by minigene reporters and exon array analysis. ChIP-seq analysis revealed that Med23 can regulate hnRNP L occupancy at their co-regulated genes. Taken together, these results demonstrate a crosstalk between Mediator and the splicing machinery, suggesting a novel mechanism for coupling mRNA processing to transcription. We performed an exon array experiment using HeLa cells expressing Med23, hnRNP L or control siRNAs which were established by a virus-mediated siRNA technology. Each sample was done in three biological replicates. Total RNA of these cell lines was processed and hybridized to the Affymetrix human exon array.

ORGANISM(S): Homo sapiens

SUBMITTER: Wencheng Li 

PROVIDER: E-GEOD-33771 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Mediator complex regulates alternative mRNA processing via the MED23 subunit.

Huang Yan Y   Li Wencheng W   Yao Xiao X   Lin Qi-Jiang QJ   Yin Jing-Wen JW   Liang Yan Y   Heiner Monika M   Tian Bin B   Hui Jingyi J   Wang Gang G  

Molecular cell 20120119 4


Mediator complex is an integrative hub for transcriptional regulation. Here we show that Mediator regulates alternative mRNA processing via its MED23 subunit. Combining tandem affinity purification and mass spectrometry, we identified a number of mRNA processing factors that bind to a soluble recombinant Mediator subunit, MED23, but not to several other Mediator components. One of these factors, hnRNP L, specifically interacts with MED23 in vitro and in vivo. Consistently, Mediator partially col  ...[more]

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