Proteomics

Dataset Information

76

Heptad-specific phosphorylation of RNA polymerase II CTD


ABSTRACT: The carboxy-terminal domain (CTD) of RNA polymerase II (Pol II) consists of heptad repeats with the consensus motif Y1-S2-P3-T4-S5-P6-S7. Dynamic phosphorylation of the CTD coordinates Pol II progression through the transcription cycle. Monoclonal antibodies have been used to study in vivo the potentially phosphorylated CTD amino acids (Y1, S2, T4, S5 and S7). However, the epitopes detected by antibodies can be masked by proteins or modifications at neighbouring sites. Therefore, the effectiveness of antibodies in western blot or ChIP analysis reflects the number of accessible CTD phosphorylation marks, but not the total number of phosphorylations. Most importantly, CTD phospho-specific antibodies do not provide any heptad - (location) specific information of CTD phosphorylation. Due to these limitations, the principles and patterns of CTD phosphorylation remained elusive. Here, we use genetic and mass spectrometric approaches to directly detect and map phosphosites along the entire CTD. We confirm phosphorylation of CTD residues Y1, S2, T4, S5 and S7 in mammalian and yeast cells. Although specific phosphorylation signatures dominate, adjacent CTD repeats can be differently phosphorylated, leading to a high variation of coexisting phosphosites in mono- and di-heptad CTD repeats. Inhibition of CDK9 kinase specifically reduces S2 phosphorylation levels within the CTD.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Mus musculus   Homo sapiens   Saccharomyces cerevisiae  

TISSUE(S): Cell Culture

DISEASE(S): Lymphoma

SUBMITTER: Ignasi Forne  

LAB HEAD: Axel Imhof

PROVIDER: PXD003159 | Pride | 2016-01-25

REPOSITORIES: Pride

altmetric image

Publications


The carboxy-terminal domain (CTD) of RNA polymerase II (Pol II) consists of heptad repeats with the consensus motif Y1-S2-P3-T4-S5-P6-S7. Dynamic phosphorylation of the CTD coordinates Pol II progression through the transcription cycle. Here, we use genetic and mass spectrometric approaches to directly detect and map phosphosites along the entire CTD. We confirm phosphorylation of CTD residues Y1, S2, T4, S5, and S7 in mammalian and yeast cells. Although specific phosphorylation signatures domin  ...[more]

Similar Datasets

2018-10-17 | PXD006004 | Pride
2015-12-22 | E-GEOD-72876 | ArrayExpress
| GSE72698 | GEO
2014-12-03 | E-GEOD-60700 | ArrayExpress
2018-10-05 | PXD008197 | Pride
2020-01-01 | E-MTAB-8522 | ArrayExpress
2012-06-04 | E-GEOD-37519 | ArrayExpress
| GSE95417 | GEO
2012-06-29 | E-MTAB-1060 | ArrayExpress
| GSE95418 | GEO