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PXD003764

PXD003764 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleTyrosine 601 of Bacillus subtilis DnaK undergoes phosphorylation and is crucial for chaperone activity and heat shock survival
DescriptionIn order to screen for cellular substrates of the Bacillus subtilis BY-kinase PtkA, and its cognate phosphotyrosine-protein phosphatase PtpZ, we performed a triple SILAC-based quantitative phosphoproteome analysis. Detected tyrosine phosphorylation sites for which the phosphorylation level decreased in the ΔptkA strain and increased in the ΔptpZ strain, compared to the wild type, were considered as potential substrates of PtkA/PtpZ. One of those sites was the residue tyrosine 601 of the molecular chaperone DnaK. We confirmed that DnaK is a substrate of PtkA and PtpZ by in vitro phosphorylation and dephosphorylation assays. In vitro, the non-phosphorylatable mutant DnaK Y601F exhibited impaired interaction with its co-chaperones DnaJ and GrpE, along with diminished capacity to hydrolyze ATP and assist the re-folding of denatured proteins. In vivo, loss of DnaK phosphorylation in the mutant strain dnaK Y601F, or in the strain overexpressing the phosphatase PtpZ, led to diminished survival upon heat shock, consistent with the in vitro results. The decreased survival of the mutant dnaK Y601F at an elevated temperature could be rescued by complementing with the wild type dnaK allele expressed ectopically. We concluded that the residue tyrosine 601 of DnaK can be phosphorylated and dephosphorylated by PtkA and PtpZ, respectively. Furthermore, Y601 is important for DnaK chaperone activity and heat shock survival of B. subtilis.
HostingRepositoryPRIDE
AnnounceDate2016-05-09
AnnouncementXMLSubmission_2016-05-09_03:26:43.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterNicolas Nalpas
SpeciesList scientific name: Bacillus subtilis; NCBI TaxID: 1423;
ModificationListphosphorylated residue
InstrumentLTQ; LTQ Orbitrap Elite
Dataset History
RevisionDatetimeStatusChangeLog Entry
02016-03-10 02:51:22ID requested
12016-05-09 03:26:45announced
Publication List
Shi L, Ravikumar V, Derouiche A, Macek B, Mijakovic I, Tyrosine 601 of Bacillus subtilis DnaK Undergoes Phosphorylation and Is Crucial for Chaperone Activity and Heat Shock Survival. Front Microbiol, 7():533(2016) [pubmed]
Keyword List
curator keyword: Biological
submitter keyword: Quantitative phosphoproteomics, bacterial protein-tyrosine kinases, molecular chaperons, protein folding, protein phosphorylation, SILAC
Contact List
Ivan Mijakovic
contact affiliationProfessor of Systems Microbiology Director of Chalmers Area of Advance Life Science Engineering Systems and Synthetic Biology Chalmers University of Technology Kemivägen 10 SE-412 96 Göteborg SWEDEN
contact emailivan.mijakovic@chalmers.se
lab head
Nicolas Nalpas
contact affiliationTuebingen University
contact emailnalpas.nicolas@gmail.com
dataset submitter
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