PXD010511

PXD010511 is an original dataset announced via ProteomeXchange.

Title	Chemical genetic identification of CDKL5 substrates reveals its role in neuronal microtubule dynamics
Description	Loss-of-function mutations in CDKL5 kinase causes severe neurodevelopmental delay and early-onset seizures. Identification of CDKL5 substrates is key to understanding its function. Using chemical genetics, we found that CDKL5 phosphorylates three microtubule-associated proteins: MAP1S, EB2 and ARHGEF2, and determined the phosphorylation sites. Substrate phosphorylations are greatly reduced in CDKL5 knockout mice, verifying these as physiological substrates. In CDKL5 knockout mouse neurons, dendritic microtubules have longer EB3-labelled plus-end growth duration and these altered dynamics are rescued by reduction of MAP1S levels through shRNA expression, indicating that CDKL5 regulates microtubule dynamics via phosphorylation of MAP1S. We show that phosphorylation by CDKL5 is required for MAP1S dissociation from microtubules. Additionally, anterograde cargo trafficking is compromised in CDKL5 knockout mouse dendrites. Finally, EB2 phosphorylation is reduced in patient-derived human neurons. Our results reveal a novel activity-dependent molecular pathway in dendritic microtubule regulation and suggest a pathological mechanism which may contribute to CDKL5 deficiency disorder.
HostingRepository	PRIDE
AnnounceDate	2018-09-18
AnnouncementXML	Submission_2018-10-02_07:33:51.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lucas Baltussen
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	phosphorylated residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2018-07-20 03:49:54	ID requested
1	2018-09-18 01:54:28	announced
⏵ 2	2018-10-02 07:33:52	announced	Updated publication reference for PubMed record(s): 30266824.

Baltussen LL, Negraes PD, Silvestre M, Claxton S, Moeskops M, Christodoulou E, Flynn HR, Snijders AP, Muotri AR, Ultanir SK, Chemical genetic identification of CDKL5 substrates reveals its role in neuronal microtubule dynamics. EMBO J, 37(24):(2018) [pubmed]

curator keyword: Biological

submitter keyword: Chemical genetics/ CDKL5/ MAP1S/ EB2/ Microtubule dynamics

Sila Ultanir
contact affiliation	The Francis Crick Institute London, UK
contact email	sila.ultanir@crick.ac.uk
lab head
Lucas Baltussen
contact affiliation	The Francis Crick Institute
contact email	lucas.baltussen@crick.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD010511
     1. Label: PRIDE project
     2. Name: Chemical genetic identification of CDKL5 substrates reveals its role in neuronal microtubule dynamics