PXD024640-1

PXD024640 is an original dataset announced via ProteomeXchange.

Title	BS3 intermolecular crosslinking map of SAGA and SLIK (SAGA-like) transcription coactivators
Description	The SAGA-like complex SLIK is a modified version of the Spt-Ada-Gcn5-Acetyltransferase (SAGA) complex. SLIK is formed through C-terminal truncation of the Spt7 SAGA subunit, causing loss of Spt8 that interacts with the TATA-binding protein. SLIK and SAGA are both coactivators of RNA polymerase II transcription in yeast. In addition, both SAGA and SLIK perform chromatin modifications and the two complexes have been speculated to uniquely contribute to transcription regulation. To test the respective contribution of SAGA vs. SLIK in transcription regulation, we assayed the chromatin modifying functions of SAGA vs. SLIK, revealing identical kinetics on minimal substrates in vitro. Furthermore, we determined a low-resolution cryo-EM structure of SLIK, revealing a modular architecture identical to SAGA. Finally, we performed a comprehensive study of DNA-binding properties of both coactivators. Purified SAGA and SLIK both associate with ssDNA and dsDNA with high affinity (KD = 10-17 nM) and the binding is sequence-independent. In conclusion, our study shows that the cleavage of Spt7 and the absence of Spt8 subunit in SLIK neither drive any major conformational differences in its structure compared to SAGA, nor significantly affect HAT, DUB or DNA binding activities in vitro.
HostingRepository	PRIDE
AnnounceDate	2021-04-19
AnnouncementXML	Submission_2021-04-18_18:45:35.063.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ralf Schittenhelm
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-03-10 03:25:46	ID requested
⏵ 1	2021-04-18 18:45:35	announced
2	2021-04-18 22:56:29	announced	2021-04-19: Updated project metadata.

10.1016/J.JBC.2021.100671;

submitter keyword: Transcriptional coactivator, Cryo-electron microscopy, Histone deubiquitination, Histone acetylation, DNA-protein interaction

Marion Boudes
contact affiliation	Biomedicine Discovery Institute, Department of Biochemistry and Molecular Biology, Monash University, Clayton VIC 3800, Australia
contact email	marion.boudes@monash.edu
lab head
Ralf Schittenhelm
contact affiliation	Monash University
contact email	ralf.schittenhelm@monash.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD024640
     1. Label: PRIDE project
     2. Name: BS3 intermolecular crosslinking map of SAGA and SLIK (SAGA-like) transcription coactivators