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Establishment of TurboID proximity proteomics in the filamentous fungus Sordaria macrospora

ABSTRACT: Biotin identification (BioID) is a proximity-dependent labeling technique to study protein-protein co-localization in vivo. Although BioID has been applied in animal cells, plants and yeast, the method remained to be established in filamentous fungi. In this study, we established BioID for the filamentous fungus Sordaria macrospora using the well-characterized striatin interacting phosphatase and kinase (STRIPAK) complex as a proof of principle. In detail, the STRIPAK complex interactor 1 (SCI1) was fused to a codon-optimized TurboID biotin ligase. This SCI1-TurboID fusion protein complemented the Δsci1 deletion strain phenotype. The identification of the already known SmSTRIPAK components PRO11, SmMOB3, SmPP2Ac1 and PRO22 in a BioID experiment with SCI1-TurboID demonstrated its successful application in S. macrospora. The technique will provide a powerful proteomics tool for fungal biologists.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Sordaria Macrospora

SUBMITTER: Oliver Valerius

LAB HEAD: Stefanie Pöggeler

PROVIDER: PXD034217 | Pride | 2022-10-25

REPOSITORIES: Pride

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Establishment of in vivo proximity labeling with biotin using TurboID in the filamentous fungus Sordaria macrospora.

Hollstein Lucas S LS Schmitt Kerstin K Valerius Oliver O Stahlhut Gertrud G Pöggeler Stefanie S

Scientific reports 20221022 1

Proximity-dependent biotin identification (BioID) has emerged as a powerful methodology to identify proteins co-localizing with a given bait protein in vivo. The approach has been established in animal cells, plants and yeast but not yet in filamentous fungi. BioID relies on promiscuous biotin ligases fused to bait proteins to covalently label neighboring proteins with biotin. Biotinylated proteins are specifically enriched through biotin affinity capture from denatured cell lysates and subseque ...[more]

PMID: 36272986

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