PXD066877

PXD066877 is an original dataset announced via ProteomeXchange.

Title	LF2 phosphorylation of the CDKL5 activation loop controls CDKL5’s activity
Description	Cdkl5 Deficiency Disorder (CDD) is caused by variants in the protein kinase CDKL5, leading to symptoms such as seizures, developmental delay, and severe intellectual disability. The Chlamydomonas homologue of human CDKL5 is the flagellar protein LF5, whose absence results in a long flagella phenotype. We found that mouse CDKL5 similarly localizes within cilia, and its loss causes long cilia. Chlamydomonas cells lacking CDKL5 exhibit altered flagellar waveforms and reduced motility. CDKL5 kinase activity is essential for flagella length control and proper localization as a kinase-dead mutant, CDKL5K33R, fails to rescue the long-flagella phenotype and does not properly localize to the proximal end of flagella. In wild-type cells, CDKL5 is highly phosphorylated at residues S162, T164, and Y166 within its activation loop and can undergo autophosphorylation in vitro. Interestingly, CDKL5 lacking kinase activity maintains similar phosphorylation at these residues. However, CDKL5 isolated from long flagella 2 cells, which lack the protein kinase LF2 shows reduced activation loop phosphorylation, diminished autophosphorylation capacity, and altered ciliary localization suggesting that LF2 phosphorylates CDKL5’s activation loop to regulate its kinase activity. Disruption of Cdk20, the mouse ortholog of LF2, likewise alters the ciliary localization of Cdkl5 in primary cilia. CDKL5 likely regulates intraflagellar transport (IFT) as the loss CDKL5 increased the ciliary abundance of IFT proteins while decreasing the phosphorylation of IFT74.
HostingRepository	PRIDE
AnnounceDate	2025-12-15
AnnouncementXML	Submission_2025-12-15_05:19:45.321.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ying Wai Lam
SpeciesList	scientific name: Chlamydomonas reinhardtii; NCBI TaxID: NEWT:3055;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion; Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2025-08-02 03:47:08	ID requested
1	2025-11-18 06:06:14	announced
⏵ 2	2025-12-15 05:19:46	announced	2025-12-15: Updated project metadata.

10.1371/journal.pbio.3003560;

Hou Y, Omi OA, Stuck MW, Cheng X, Walker B, Lam YW, Schmoker AM, Nguyen SN, Gonzalez-Perez MP, Ballif BA, Lechtreck KF, Witman GB, Pazour GJ, Activation of the ciliary kinase CDKL5 is mediated by the cyclin-dependent kinase CDK20/LF2 to control flagellar length. PLoS Biol, 23(12):e3003560(2025) [pubmed]

submitter keyword: Chlamydomonas, cilia,Cdkl5 Deficiency Disorder, ciliopathy, intraflagellar transport, kinase, flagella

Gregory J
contact affiliation	Program in Molecular Medicine, UMass Chan Medical School
contact email	Gregory.Pazour@umassmed.edu
lab head
Ying Wai Lam
contact affiliation	Vermont Biomedical Research Network Proteomics Facility Department of Biology University of Vermont
contact email	ylam@uvm.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/11/PXD066877

PRIDE project URI

• PRIDE
  1. PXD066877
     1. Label: PRIDE project
     2. Name: LF2 phosphorylation of the CDKL5 activation loop controls CDKL5’s activity