PXD015963-1

PXD015963 is an original dataset announced via ProteomeXchange.

Title	Comparative phosphorylation map of Dishevelled 3 links phospho-signatures to biological outputs.
Description	Background: Dishevelled (DVL) is an essential component of the Wnt signaling cascades. Function of DVL is controlled by phosphorylation but the molecular details are missing. DVL3 contains 131 serines and threonines whose phosphorylation generates complex barcodes underlying diverse DVL3 functions. In order to dissect the role of DVL phosphorylation we analyzed the phosphorylation of human DVL3 induced by previously reported (CK1ε, NEK2, PLK1, CK2α, RIPK4, PKCδ) and newly identified (TTBK2, Aurora A) DVL kinases. Methods: Shotgun proteomics including TiO2 enrichment of phosphorylated peptides followed by liquid chromatography tandem mass spectrometry on immunoprecipitates from HEK293T cells was used to identify and quantify phosphorylation of DVL3 protein induced by 8 kinases. Functional characterization was performed by in-cell analysis of phospho-mimicking/non phosphorylatable DVL3 mutants and supported by FRET assays and NMR spectroscopy. Results: We used quantitative mass spectrometry and calculated site occupancies and quantified phosphorylation of >80 residues. Functional validation demonstrated the importance of CK1ε-induced phosphorylation of S268 and S311 for Wnt-3a-induced β-catenin activation. S630-643 cluster phosphorylation by CK1, NEK2 or TTBK2 is essential for even subcellular distribution of DVL3 when induced by CK1 and TTBK2 but not by NEK2. Further investigation showed that NEK2 utilizes a different mechanism to promote even localization of DVL3. NEK2 triggered phosphorylation of PDZ domain at S263 and S280 prevents binding of DVL terminus to PDZ and promotes an open conformation of DVL3 that is more prone to even subcellular localization. Conclusions: We identify unique phosphorylation barcodes associated with DVL function. Our data provide an example of functional synergy between phosphorylation in structured domains and unstructured IDRs that together dictate the biological outcome.
HostingRepository	PRIDE
AnnounceDate	2020-01-09
AnnouncementXML	Submission_2020-01-09_04:32:39.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Katerina Hanáková
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2019-10-22 06:31:34	ID requested
⏵ 1	2020-01-09 04:32:40	announced
2	2024-10-22 03:59:04	announced	2024-10-22: Updated project metadata.

Han, á, kov, á K, Bernat, í, k O, Kravec M, Micka M, Kumar J, Harno, š J, Ovesn, á P, Pacl, í, kov, á P, R, á, dsetoulal M, Pot, ě, š, il D, Tripsianes K, Č, aj, á, nek L, Zdr, á, hal Z, Bryja V, Comparative phosphorylation map of Dishevelled 3 links phospho-signatures to biological outputs. Cell Commun Signal, 17(1):170(2019) [pubmed]

submitter keyword: Dishevelled/DVL3/phosphorylation/kinase/mass spectrometry/CK1/TTBK2/NEK2/Wnt

Zbynek Zdrahal
contact affiliation	CEITEC—Central European Institute of Technology, Masaryk University, Brno, Czech Republic National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, Czech Republic
contact email	zdrahal@sci.muni.cz
lab head
Katerina Hanáková
contact affiliation	CEITEC MU
contact email	katerina.hanak@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD015963
     1. Label: PRIDE project
     2. Name: Comparative phosphorylation map of Dishevelled 3 links phospho-signatures to biological outputs.