PXD006067

PXD006067 is an original dataset announced via ProteomeXchange.

Title	A mitogen-activated protein kinase limits the apical formation of phosphatidylinositol 4,5-bisphosphate during polarized cell expansion of pollen tubes
Description	Mitogen activated protein kinases (MAPKs) and phosphoinositides contribute to establishment and maintenance of polarity in eukaryotic cells. However, interactions between these important signaling pathways are largely unclear. Pollen tubes are an important model for the study of polarized cell expansion, in which PI4P 5-kinases control apical secretion and MAPKs contribute to pollen tube guidance. Here we report on the regulation of the Arabidopsis PI4P 5-kinase, AtPIP5K6, by the MAPK, MPK6. Purified recombinant AtPIP5K6 was phosphorylated in vitro upon incubation with pollen tube extracts. Non-targeted in-gel-kinase assays of pollen tube extracts followed by mass-spectrometry identified MPK6 as a candidate protein kinase upstream of AtPIP5K6. MPK6 interacted with AtPIP5K6 in yeast-two-hybrid tests, and bimolecular fluorescence complementation verified the interaction at the apical plasma membrane of pollen tubes. Recombinant AtPIP5K6 was phosphorylated in vitro by recombinant MPK6, resulting in reduced catalytic activity of AtPIP5K6. Alanine-substitution of phosphorylation sites, T590A and T597A, abrogated phosphorylation of AtPIP5K6 by MPK6 and prevented the in vitro inhibition. A T597D substitution resulted in reduced catalytic activity of the modified AtPIP5K6 protein in vitro. Coexpression of MPK6 with AtPIP5K6 in pollen tubes resulted in attenuation of PI4P 5-kinase-mediated polarity defects in vivo. Reciprocally, AtPIP5K6 effects were enhanced upon concomitant RNAi-inhibition of endogenous tobacco MAPKs. Both effects were absent when a AtPIP5K6 T590A T597A-variant was expressed, which could no longer be phosphorylated. We conclude that MPK6 limits PI4P 5-kinase-dependent polarized secretion to effect pollen tube guidance, revealing a new link between conserved signaling pathways with potential ramifications across eukaryotic kingdoms.
HostingRepository	PRIDE
AnnounceDate	2022-02-22
AnnouncementXML	Submission_2022-02-22_03:46:16.205.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sacha Baginsky
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702; scientific name: Nicotiana tabacum (Common tobacco); NCBI TaxID: 4097;
ModificationList	phosphorylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Velos; Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2017-03-10 05:48:37	ID requested
⏵ 1	2022-02-22 03:46:17	announced

Hempel F, Stenzel I, Heilmann M, Krishnamoorthy P, Menzel W, Golbik R, Helm S, Dobritzsch D, Baginsky S, Lee J, Hoehenwarter W, Heilmann I, MAPKs Influence Pollen Tube Growth by Controlling the Formation of Phosphatidylinositol 4,5-Bisphosphate in an Apical Plasma Membrane Domain. Plant Cell, 29(12):3030-3050(2017) [pubmed]

curator keyword: Biological

submitter keyword: Polar tip growth, MAP-kinase, PI4P 5-kinase, phosphorylation, PtdIns(4,5)P2

Ingo Heilmann
contact affiliation	Department of Cellular Biochemistry Institute for Biochemistry and Biotechnology Martin-Luther-University Halle-Wittenberg Kurt-Mothes-Str. 3 06120 Halle (Saale)
contact email	ingo.heilmann@biochemtech.uni-halle.de
lab head
Sacha Baginsky
contact affiliation	Institut fuer Biochemie und Biotechnologie
contact email	pflanzen@biochemtech.uni-halle.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD006067
     1. Label: PRIDE project
     2. Name: A mitogen-activated protein kinase limits the apical formation of phosphatidylinositol 4,5-bisphosphate during polarized cell expansion of pollen tubes