PXD069539

PXD069539 is an original dataset announced via ProteomeXchange.

Title	Proxiomes of the greenbeard proteins SmDOC1 and SmDOC2 in the filamentous fungus Sordaria macrospora
Description	In this study, we used the in vivo proximity labeling biotin identification (BioID; Roux et al., 2012) method to capture proteins of the microenvironment of the greenbeard determinant of communication (DOC) proteins SmDOC1 and SmDOC2 in the filamentous fungus Sordaria macrospora. The genes Smdoc1 (SMAC_06903) and Smdoc2 (SMAC_06902) were fused to the TurboID ligase via a GGGGS x 2 Linker (Branon et al., 2018) and integrated at their native chromosomal locus. To ensure authentic expression regulation, the Smdoc-TurboID fusion genes were under the control of their native 5’ regions. The homologous doc genes of the closely related fungus Neurospora crassa were shown to be involved in long distance kind discrimination, regulation hyphal fusion upon compatible allorecognition (Heller et al., 2016). The S. macrospora Smdoc1/2 are required for proper sexual development. Our SmDOC1-BioID experimental setup comprises five biological replicates of the strain expressing the Smdoc1-TurboID fusion gene from its native locus and a control strain expressing an egfp-TurboID fusion gene under control of the 1 kb 5’ Smdoc1 promoter region, which was ectopically integrated into the S. macrospora wild type. Our SmDOC2-BioID experimental setup consists of four biological replicates of three strains: 1) a strain expressing the free egfp-TurboID fusion gene under control of the 1 kb 5’ Smdoc1 promoter region, ectopically integrated into the S. macrospora wild type; 2) a strain expressing the Smdoc2-TurboID fusion gene from its native locus; 3) a strain expressing the Smdoc2-TurboID fusion gene from its native locus in the Δmak2 deletion strain background which lacks the MAP kinase mak2 of the pheromone response pathway (Schmidt et al., 2020). To account for possible expression artefacts among strains used in the SmDOC1- and SmDOC2-BioID experiments, we performed additional input controls of the crude protein extract prior to biotin affinity purification. Roux, K. J., Kim, D. in, Raida, M., & Burke, B. (2012). A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. The Journal of Cell Biology, 196(6), 801–810. Branon, T. C., Bosch, J. A., Sanchez, A. D., Udeshi, N. D., Svinkina, T., Carr, S. A., Feldman, J. L., Perrimon, N., & Ting, A. Y. (2018). Efficient proximity labeling in living cells and organisms with TurboID. Nature Biotechnology, 36(9), 880–887. Heller, J., Zhao, J., Rosenfield, G., Kowbel, D. J., Gladieux, P., & Glass, N. L. (2016). Characterization of Greenbeard Genes Involved in Long-Distance Kind Discrimination in a Microbial Eukaryote. PLOS Biology, 14(4), e1002431. Schmidt, S., Märker, R., Ramšak, B., Beier-Rosberger, A. M., Teichert, I., & Kück, U. (2020). Crosstalk Between Pheromone Signaling and NADPH Oxidase Complexes Coordinates Fungal Developmental Processes. Frontiers in Microbiology, 11, 1722.
HostingRepository	PRIDE
AnnounceDate	2026-08-19
AnnouncementXML	Submission_2026-08-19_04:17:28.334.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Oliver Valerius
SpeciesList	scientific name: Sordaria macrospora; NCBI TaxID: NEWT:5147;
ModificationList	biotinylated residue; phosphorylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2025-10-16 11:57:47	ID requested
⏵ 1	2026-08-19 04:17:28	announced

10.1371/JOURNAL.PBIO.3003969;

submitter keyword: Sordaria macrospora, TurboID, proximity labeling, BioID, determinant of communication

Dr. Prof. Stefanie Pöggeler
contact affiliation	Georg-August Universität Göttingen, 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

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PRIDE project URI

• PRIDE
  1. PXD069539
     1. Label: PRIDE project
     2. Name: Proxiomes of the greenbeard proteins SmDOC1 and SmDOC2 in the filamentous fungus Sordaria macrospora