PXD068782-1

PXD068782 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 cells that lack the protein kinase LF2 has 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	2025-11-15
AnnouncementXML	Submission_2025-11-15_06:24:39.145.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Gregory Pazour
SpeciesList	scientific name: Chlamydomonas reinhardtii; NCBI TaxID: NEWT:3055;
ModificationList	phosphorylated residue
Instrument	timsTOF Pro 2

Revision	Datetime	Status	ChangeLog Entry
0	2025-09-24 13:06:11	ID requested
⏵ 1	2025-11-15 06:24:39	announced
2	2025-12-15 05:20:25	announced	2025-12-15: Updated project metadata.

Dataset with its publication pending

submitter keyword: Cilia,CDKL5, phospho-proteome, Flagella, CDD, Chlamydomonas, Cdk20, LF5, activation loop, LF2, phosphorylation, kinase, proteome, Cdkl5 deficiency disorder

Gregory Pazour
contact affiliation	Program in Molecular Medicine UMass Chan Medical School Worcester MA 01605 USA
contact email	gregory.pazour@umassmed.edu
lab head
Gregory Pazour
contact affiliation	University of Massachusetts Chan Medical School
contact email	gregory.pazour@umassmed.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD068782
     1. Label: PRIDE project
     2. Name: LF2 phosphorylation of the CDKL5 activation loop controls CDKL5’s activity