PXD038688

PXD038688 is an original dataset announced via ProteomeXchange.

Title	B4 Raf-like MAPKKK RAF24 regulates Arabidopsis thaliana flowering time
Description	The timing of flowering is a critical agronomic trait governed by a number of different internal and external events, such as temperature, light and photoperiod. Despite a firm understanding of flowering time from a genetic standpoint, there exists a limited understanding of how post-translational modifications may influence flowering, and what molecular PTM machinery is responsible for catalyzing these events. Here, we identify and characterize a novel B4 Raf-like MAPKKK protein kinase called RAF24, which exhibits an early flowering phenotype. Analysis of closely-related B4 Raf-like MAPKKK protein kinases RAF18 and RAF20, indicate that RAF24 is the only B4 RAF to possess an early flowering phenotype. Through quantitative proteomics, we then found that RAF24 impacts the phosphorylation status of numerous flowering related proteins related to the autonomous flowering pathway, with changes in the phosphorylation status of HISTONE MONO-UBIQUITINATION 2 (HUB2) representing the largest fold-change. Subsequent analysis of the HUB2 protein interactome by quantitative affinity purification mass spectrometry (AP-MS) revealed a relationship between HUB2 phosphorylation at S314 phosphorylation status and HUB2 function that manifest as changes in activity and protein-protein interactions. Use of phospho-memetic and -ablative mutants validate the importance of S314, directly linking this HUB2 phosphorylation event to the proper control of flowering time. Collectively, our findings implicate RAF24 as a higher-order regulator of flowering, while further implicating HUB2 as a centerpiece of flowering time regulation in plants.
HostingRepository	PRIDE
AnnounceDate	2025-05-09
AnnouncementXML	Submission_2025-05-08_20:52:39.628.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Richard Uhrig
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	phosphorylated residue
Instrument	Orbitrap Fusion Lumos; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2022-12-08 16:09:50	ID requested
⏵ 1	2025-05-08 20:52:40	announced

Dataset with its publication pending

submitter keyword: Phosphoproteomics, Tandem-Affinity Purification, Protein Kinases, DIA,Arabidopsis, Fllowering

Richard Glen Uhrig
contact affiliation	Department of Biological Sciences - University of Alberta
contact email	ruhrig@ualberta.ca
lab head
Richard Uhrig
contact affiliation	University of Alberta
contact email	ruhrig@ualberta.ca
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD038688
     1. Label: PRIDE project
     2. Name: B4 Raf-like MAPKKK RAF24 regulates Arabidopsis thaliana flowering time