PXD005967-1
PXD005967 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | MudPIT analysis of proteins interacting with Saccharomyces cerevisiae RNA Polymerase II HA-Rpb1 C-Terminal Domain |
| Description | The carboxy-terminal domain (CTD) of Rpb1, the largest component of the 12-subunit RNA polymerase II, consists of repeating Y1S2P3T4S5P6S7 heptapeptides (26 repeats in budding yeast). Each stage of transcription relies on the ordered recruitment and exchange of specific protein complexes that act on RNA polymerase II, its nascent transcripts, and the underlying chromatin. This dynamic process is orchestrated via patterned post-translational modifications of the CTD. To characterize the role of phosphorylation on Thr4, we examined the effect of Rpb1 alleles in which Thr4 was substituted with an alanine (T4A) or the phospho-mimic glutamate (T4E). Substitutions were made across all heptad repeats of the CTD. We affinity purified HA-tagged Rpb1 from Saccharomyces cerevisiae strains bearing WT, T4A, and T4E CTDs. A control strain (Z26) lacking the HA-tagged Rpb1 was subjected to an identical affinity enrichment procedure. Three biological replicates were acquired for each type of affinity purification and analyzed independently. After TCA-precipitation, proteins were urea-denatured, reduced, alkylated, then digested with endoproteinase LysC followed by trypsin. The resulting peptide mixtures were analyzed by Multidimensional Protein Identification Technology (MudPIT). Label-free quantitative proteomics was used to identify and quantify the relative abundance of affinity-enriched complexes. |
| HostingRepository | MassIVE |
| AnnounceDate | 2017-05-02 |
| AnnouncementXML | Submission_2017-05-02_11:17:49.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Unsupported dataset by repository |
| PrimarySubmitter | Laurence Florens |
| SpeciesList | scientific name: Saccharomyces cerevisiae; common name: baker's yeast; NCBI TaxID: 4932; |
| ModificationList | L-methionine sulfoxide; S-carboxamidomethyl-L-cysteine |
| Instrument | LTQ |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2017-02-24 15:12:48 | ID requested | |
| ⏵ 1 | 2017-05-02 11:17:50 | announced |
Publication List
| Corey M. Nemec, Fan Yang, Joshua M. Gilmore, Corinna Hintermair, Yi-Hsuan Ho, Sandra C. Tseng, Martin Heidemann, Ying Zhang, Laurence Florens, Audrey P. Gasch, Dirk Eick, Michael P. Washburn, Gabriele Varani, and Aseem Z. Ansari. Different phosphoisoforms of RNA polymerase II engage the Rtt103 termination factor in a structurally analogous manner. PNAS 2017 |
| published ahead of print May 2, 2017, doi:10.1073/pnas.1700128114. |
Keyword List
| submitter keyword: RNA polymerase II, CTD phosphorylation, carboxyl-terminal domain (CTD), protein recruitment |
Contact List
| Laurence Florens | |
|---|---|
| contact affiliation | The Stowers Institute for Medical Research |
| contact email | laf@stowers.org |
| lab head | |
| Laurence Florens | |
| contact affiliation | Stowers Institute for Medical Research |
| contact email | laf@stowers.org |
| dataset submitter | |
Full Dataset Link List
| MassIVE dataset URI |
| 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://massive.ucsd.edu/MSV000080573 |
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