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Series GSE87767 Query DataSets for GSE87767
Status Public on May 01, 2018
Title mRNA cap methyltransferase, RNMT-RAM, promotes RNA pol II transcription
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Genome binding/occupancy profiling by high throughput sequencing
Summary mRNA cap addition occurs early during RNA pol II transcription, facilitating pre-mRNA processing and translation. We report that the mammalian mRNA cap methyltransferase, RNMT-RAM, promotes RNA pol II transcription, independently of mRNA capping and translation. In cells, sub-lethal suppression of RNMT-RAM reduces RNA pol II occupancy, net mRNA synthesis and pre-mRNA levels. Conversely, expression of RNMT-RAM increases transcription independently of cap methyltransferase activity. In isolated nuclei, recombinant RNMT-RAM stimulates transcriptional output; this requires the RAM RNA-binding domain. RNMT-RAM interacts with nascent transcripts along their entire length and with transcription associated factors including RNA pol II subunits, SPT4, SPT6 and PAFc. Suppression of RNMT-RAM inhibits transcriptional markers including histone H2B K120 ubiquitination, H3 K4 and K36 methylation, RNA pol II S5 and S2 phosphorylation and PAFc recruitment. These findings suggest that multiple interactions between RNMT-RAM, RNA pol II factors and RNA along the transcription unit stimulate transcription.
Overall design CLIP experiment performed on HA-RNMT expressing HeLa cells. RNASeq and ChIP-Seq performed on HeLa cells treated with siRNA for RAM and scrambled control.
Contributor(s) Varshney D, Cowling VH
Citation(s) 29719263
Submission date Oct 07, 2016
Last update date May 15, 2019
Contact name Dhaval Varshney
Organization name University of Dundee
Department Centre for Gene Regulation and Expression
Street address Dow Street
City Dundee
ZIP/Postal code DD1 5EH
Country United Kingdom
Platforms (2)
GPL15520 Illumina MiSeq (Homo sapiens)
GPL18573 Illumina NextSeq 500 (Homo sapiens)
Samples (20)
GSM2340020 CLIP_Input
GSM2340021 CLIP_RNMT_1
GSM2340022 CLIP_RNMT_2
BioProject PRJNA347477
SRA SRP091113

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Supplementary file Size Download File type/resource
GSE87767_CLIP-counts.csv.gz 273.8 Kb (ftp)(http) CSV
GSE87767_ChIPSeq_scsiRNA.bigWig 1.3 Gb (ftp)(http) BIGWIG
GSE87767_ChIPSeq_siRAM.bigWig 1.4 Gb (ftp)(http) BIGWIG
GSE87767_RNASeq-counts.csv.gz 570.5 Kb (ftp)(http) CSV
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Processed data are available on Series record

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