Unknown,Transcriptomics,Genomics,Proteomics

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Using GRO-seq experiments to determine the mechanism of C. elegans dosage compensation's control of transcription


ABSTRACT: Dosage Compensation is required to correct for uneven gene dose between the sexes. We utilized global run-on sequencing (GRO-seq) to examine how Caenorhabditis elegans dosage compensation reduces transcription of X-linked genes. To facilitate these experiments, we required accurate 5M-bM-^@M-^Y-ends of genes that have been missing due to a co-transcriptional trans-splicing event common in nematodes. We developed a modified GRO-seq protocol to identify TSSs that are supported by transcription, and determined that TSSs lie more than 1 kb upstream of the previously annotated TSS for nearly one-quarter of all genes. We then investigated the changes that occur in transcriptionally engaged RNA Polymerase when dosage compensation is disrupted, and find that dosage compensation controls recruitment of RNA Polymerase to X-linked genes. GRO-seq experiments (two biological replicates) were performed in nuclei from many wild-type states and a dosage compensation mutant

ORGANISM(S): Caenorhabditis elegans

SUBMITTER: Barbara Meyer 

PROVIDER: E-GEOD-43084 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Condensin controls recruitment of RNA polymerase II to achieve nematode X-chromosome dosage compensation.

Kruesi William S WS   Core Leighton J LJ   Waters Colin T CT   Lis John T JT   Meyer Barbara J BJ  

eLife 20130618


The X-chromosome gene regulatory process called dosage compensation ensures that males (1X) and females (2X) express equal levels of X-chromosome transcripts. The mechanism in Caenorhabditis elegans has been elusive due to improperly annotated transcription start sites (TSSs). Here we define TSSs and the distribution of transcriptionally engaged RNA polymerase II (Pol II) genome-wide in wild-type and dosage-compensation-defective animals to dissect this regulatory mechanism. Our TSS-mapping stra  ...[more]

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