PXD061752

PXD061752 is an original dataset announced via ProteomeXchange.

Title	In vivo proximity-labeling with miniTurboID to screen for protein-protein interactions in the filamentous ascomycete Sordaria macrospora
Description	The Biotin Identification (BioID) method is an in vivo labeling strategy for the identification of cellular microenvironments in animal cells, plants, yeast and filamentous fungi. With this method, the protein of interest (POI) is fused to a promiscuous biotin ligase. The ligase biotinylates proximal proteins in a cumulative manner, which are then captured from cell lysates by affinity-binding and identified by mass spectrometry. This allows the identification of POI co-localizing proteins, including weakly or transiently binding ones (Roux et al., 2012; Branon et al., 2018). In our recent research, we performed BioID experiments with the TurboID biotin ligase expressed in the filamentous fungus Sordaria macrospora (Hollstein et al., 2022). Here, we tested a smaller variant of TurboID biotin ligase called miniTurboID for its application S. macrospora. For a proof of principle, we used the well-characterized striatin interacting phosphatase and kinase (STRIPAK) complex as molecular microenvironment. In detail, the STRIPAK complex interactor 1 (SCI1) was fused to the codon-optimized TurboID or the miniTurboID biotin ligase. These fusion proteins complemented a sterile Δsci1 deletion strain to restore fertility. A BioID experiment with LC-MS analysis showed significant enrichment of the SmSTRIPAK components PRO11, SmMOB3 and PRO22 in all biological replicates of SCI1-TurboID and SCI1-miniTurboID, while being absent in the free TurboID control. This demonstrates that both, TurboID and the smaller variant miniTurboID, can be applied in the filamentous ascomycete S. macrospora. We provide a step by step protocol for the preparation of BioID samples for LC-MS analysis in the MethodsX article.
HostingRepository	PRIDE
AnnounceDate	2025-05-12
AnnouncementXML	Submission_2025-05-12_07:42:51.789.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Oliver Valerius
SpeciesList	scientific name: Sordaria macrospora; NCBI TaxID: 5147;
ModificationList	biotinylated residue; phosphorylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2025-03-12 03:29:40	ID requested
⏵ 1	2025-05-12 07:42:52	announced

10.1016/J.MEX.2025.103351;

submitter keyword: Sordaria macrospora, STRIPAK, proximity labeling, miniTurbo, miniTurboID, BioID

Prof. Stefanie Pöggeler
contact affiliation	Georg-August University, Institute of Microbiology and Genetics, Department of Genetics of Eukaryotic Microorganisms
contact email	spoegge@gwdg.de
lab head
Oliver Valerius
contact affiliation	Georg-August-Universität Göttingen
contact email	ovaleri@gwdg.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/2025/05/PXD061752

PRIDE project URI

• PRIDE
  1. PXD061752
     1. Label: PRIDE project
     2. Name: In vivo proximity-labeling with miniTurboID to screen for protein-protein interactions in the filamentous ascomycete Sordaria macrospora