⮝ Full datasets listing
PXD073867
PXD073867 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Characterization of proteins assembled in vitro on yeast centromeric DNA in WT vs cbf1Delta extracts |
| Description | This experiment aims to characterize the proteins assembled in vitro on yeast centromeric DNA in WT (SBY3) vs cbf1Delta (SBY4958) extracts. Briefly, yeast centromeric DNA (CEN3-WT and CEN3-CDEIm) is PCR amplified, biotinylated, and immobilized to streptavidin-coated beads (M280, Invitrogen). Yeast lysate is prepared by harvesting benomyl-treated (30 mg/mL, 3 hours) yeast cultures, resuspending in Buffer L 0.175 (25 mM HEPES pH 7.6, 2 mM MgCl2, 0.1 mM EDTA pH 7.6, 0.5 mM EGTA pH 7.6, 0.1 % NP-40, 175 mM K-Glutamate, and 15% Glycerol) supplemented with protease inhibitors, and lysed using a Freezer Mill (SPEX, Metuchen NJ). Lysate was clarified via centrifugation at 13,000 x g for 30 minutes, and the protein layer was collected. The DNA on beads (60 ul) is incubated with 1500 ul yeast whole cell extracts for 90 minutes at room temperature. Then beads are washed 5 times with buffer Buffer L 0.175, and twice with 50 mM HEPES pH 8, and eluted with 60 ul 0.2% RapiGest (Waters Corporation, Milford MA) in 50mM HEPES pH 8 for 30 minutes at room temperature. SH061818_070118_SBY3_WT: lysate from WT cells (SBY3) assembled on CEN3-WT DNA template SH061818_070118_SBY3_CDEI: lysate from WT cells (SBY3) assembled on CEN3-CDEIm DNA template. This DNA template has mutations on the CDEI element which disrupt Cbf1 binding. SH061818_070118_SBY4958_WT: lysate from cbf1Delta cells (SBY4958) assembled on CEN3-WT DNA template |
| HostingRepository | MassIVE |
| AnnounceDate | 2026-03-24 |
| AnnouncementXML | Submission_2026-03-24_13:44:16.175.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Non peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Supported dataset by repository |
| PrimarySubmitter | Fred Hutch Proteomics Core |
| SpeciesList | scientific name: Saccharomyces cerevisiae; common name: baker's yeast; NCBI TaxID: 4932; |
| ModificationList | No PTMs are included in the dataset |
| Instrument | Orbitrap Fusion |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2026-01-30 15:29:28 | ID requested | |
| ⏵ 1 | 2026-03-24 13:44:16 | announced |
Publication List
| no publication |
Keyword List
| submitter keyword: Key words: Mitosis, Kinetochore, Cbf1, CCAN, DatasetType:Proteomics |
Contact List
| Susan Biggins | |
|---|---|
| contact affiliation | Fred Hutchinson Cancer Center |
| contact email | sbiggins@fredhutch.org |
| lab head | |
| Fred Hutch Proteomics Core | |
| contact affiliation | Fred Hutchinson Cancer Research Center |
| contact email | pgafken@fredhutch.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-ftp.ucsd.edu/v12/MSV000100678/ |
