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| 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
Other
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| 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.
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| 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.
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| Contributor(s) |
Varshney D, Cowling VH |
| Citation(s) |
29719263 |
| Submission date |
Oct 07, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Dhaval Varshney |
| E-mail(s) |
d.varshney@dundee.ac.uk
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| Organization name |
University of Dundee
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| Department |
Centre for Gene Regulation and Expression
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| Street address |
Dow Street
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| City |
Dundee |
| ZIP/Postal code |
DD1 5EH |
| Country |
United Kingdom |
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| Platforms (2) |
| GPL15520 |
Illumina MiSeq (Homo sapiens) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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| Samples (20)
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| Relations |
| BioProject |
PRJNA347477 |
| SRA |
SRP091113 |
| 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 |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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