PXD037249

PXD037249 is an original dataset announced via ProteomeXchange.

Title	A SAM key domain required for enzymatic activity and allosteric activation of the Fun30 nucleosome remodeler
Description	Fun30 is the prototype of the Fun30-SMARCAD1-ETL sub-family of nucleosome remodelers involved in DNA repair and gene silencing. These proteins appear to act as single subunit nucleosome remodelers, but their molecular mechanisms are, at this point, poorly understood. Using multiple sequence alignment and structure prediction, we identify an evolutionarily conserved domain that is modeled to contain a SAM-like fold with one long, protruding helix, which we term SAM-key. Deletion of the SAM-key within budding yeast Fun30 leads to a defect in DNA repair and gene silencing similar to that of the fun30 mutant. In vitro, Fun30 protein lacking the SAM key is able to bind nucleosomes but is deficient in DNA-stimulated ATPase activity as well as nucleosome sliding and eviction. A structural model based on AlphaFold2 prediction and verified by crosslinking-MS indicates an interaction of the long SAM-key helix with protrusion I, a subdomain located between the two ATPase lobes that is critical for control of enzymatic activity. Mutation of the interaction interface phenocopies the domain deletion with a lack of DNA-stimulated ATPase activation and a nucleosome remodeling defect, thereby confirming a role of the SAM-key helix in regulating ATPase activity. Our data thereby demonstrate a central role of the SAM-key domain in mediating the activation of Fun30 catalytic activity, thus highlighting the importance of allosteric activation for this class of enzymes.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_07:13:28.702.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Barbara Steigenberger
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2022-10-10 07:42:37	ID requested
1	2023-07-18 01:11:33	announced
⏵ 2	2023-11-14 07:13:29	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: SAM, Fun30,yeast, chromatin, resection, regulation, key

Boris Pfander
contact affiliation	Institute of Aerospace Medicine German Aerospace Center (DLR) Linder Höhe 51147 Cologne, Germany & Institute for Genome Stability in Ageing and Disease University of Cologne CECAD Research Center Joseph-Stelzmann-Str. 26 50931 Cologne, Germany & Max-Planck Institute of Biochemistry Am Klopferspitz 18 82152 Martinsried, Germany
contact email	bpfander@biochem.mpg.de
lab head
Barbara Steigenberger
contact affiliation	MPI of Biochemistry
contact email	steigenberger@biochem.mpg.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/2023/07/PXD037249

PRIDE project URI

• PRIDE
  1. PXD037249
     1. Label: PRIDE project
     2. Name: A SAM key domain required for enzymatic activity and allosteric activation of the Fun30 nucleosome remodeler