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High-Throughput Genetic and Gene Expression Analysis of the RNAPII-CTD Reveals Unexpected Connections to SRB10/CDK8
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High-Throughput Genetic and Gene Expression Analysis of the RNAPII-CTD Reveals Unexpected Connections to SRB10/CDK8
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High-Throughput Genetic and Gene Expression Analysis of the RNAPII-CTD Reveals Unexpected Connections to SRB10/CDK8
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Journal Article
High-Throughput Genetic and Gene Expression Analysis of the RNAPII-CTD Reveals Unexpected Connections to SRB10/CDK8
2013
Overview
The C-terminal domain (CTD) of RNA polymerase II (RNAPII) is composed of heptapeptide repeats, which play a key regulatory role in gene expression. Using genetic interaction, chromatin immunoprecipitation followed by microarrays (ChIP-on-chip) and mRNA expression analysis, we found that truncating the CTD resulted in distinct changes to cellular function. Truncating the CTD altered RNAPII occupancy, leading to not only decreases, but also increases in mRNA levels. The latter were largely mediated by promoter elements and in part were linked to the transcription factor Rpn4. The mediator subunit Cdk8 was enriched at promoters of these genes, and its removal not only restored normal mRNA and RNAPII occupancy levels, but also reduced the abnormally high cellular amounts of Rpn4. This suggested a positive role of Cdk8 in relationship to RNAPII, which contrasted with the observed negative role at the activated INO1 gene. Here, loss of CDK8 suppressed the reduced mRNA expression and RNAPII occupancy levels of CTD truncation mutants.
Publisher
Public Library of Science,Public Library of Science (PLoS)
Subject
/ Chromatin Immunoprecipitation
/ Cyclin-Dependent Kinase 8 - genetics
/ DNA-Binding Proteins - genetics
/ DNA-Binding Proteins - metabolism
/ Gene Expression Regulation, Fungal
/ Kinases
/ Mutation
/ Myo-Inositol-1-Phosphate Synthase - genetics
/ Myo-Inositol-1-Phosphate Synthase - metabolism
/ Ontology
/ Plasmids
/ RNA Polymerase II - chemistry
/ RNA Polymerase II - genetics
/ Saccharomyces cerevisiae - genetics
/ Saccharomyces cerevisiae Proteins - genetics
/ Saccharomyces cerevisiae Proteins - metabolism
/ Transcription Factors - genetics
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