PXD003159

PXD003159 is an original dataset announced via ProteomeXchange.

Title	Heptad-specific phosphorylation of RNA polymerase II CTD
Description	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.
HostingRepository	PRIDE
AnnounceDate	2016-01-25
AnnouncementXML	Submission_2016-01-25_03:11:08.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ignasi Forne
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	iodoacetamide derivatized residue; phosphorylated residue
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2015-11-06 07:43:04	ID requested
⏵ 1	2016-01-25 03:11:10	announced

Sch, ü, ller R, Forn, é I, Straub T, Schreieck A, Texier Y, Shah N, Decker TM, Cramer P, Imhof A, Eick D, Heptad-Specific Phosphorylation of RNA Polymerase II CTD. Mol Cell, 61(2):305-14(2016) [pubmed]

curator keyword: Biological

submitter keyword: Pol II CTD

CTD heptads

CTD phosphosites

LC-MS/MS

Axel Imhof
contact affiliation	Biomedical Center Munich Molecular Biology Großhaderner Strasse 9 82152 Planegg-Martinsried
contact email	imhof@lmu.de
lab head
Ignasi Forne
contact affiliation	Biomedical Center-LMU
contact email	ignasi.forne@lrz.uni-muenchen.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/01/PXD003159

PRIDE project URI

• PRIDE
  1. PXD003159
     1. Label: PRIDE project
     2. Name: Heptad-specific phosphorylation of RNA polymerase II CTD