PXD048637

PXD048637 is an original dataset announced via ProteomeXchange.

Title	CAMSAP-driven microtubule release from γ-TuRC and its regulation by nucleation-promoting factors
Description	γ-tubulin ring complex (γ-TuRC) is the major microtubule-nucleating factor. After nucleation, microtubules can be released from γ-TuRC and stabilized by other proteins, such as CAMSAPs, but the biochemical cross-talk between minus-end regulation pathways is poorly understood. Here, we reconstituted this process in vitro using purified components. We found that all CAMSAPs could bind to the minus-ends of γ-TuRC-attached microtubules. CAMSAP2 and CAMSAP3, which decorate and stabilize growing minus ends, but not the minus-end tracking protein CAMSAP1 induced microtubule release from γ-TuRC. CDK5RAP2, a γ-TuRC-interactor, and CLASP2, a regulator of microtubule growth, strongly stimulated γ-TuRC-dependent microtubule nucleation, but only CDK5RAP2 suppressed CAMSAP binding to γ-TuRC-anchored minus ends and their release. CDK5RAP2 also improved selectivity of γ-tubulin-containing complexes for 13- rather than 14-protofilament microtubules in microtubule-capping assays. Knockout and overexpression experiments in cells showed that CDK5RAP2 inhibits formation of CAMSAP2-bound microtubules detached from the microtubule-organizing center. We conclude that CAMSAPs can release newly nucleated microtubules from γ-TuRC, whereas nucleation-promoting factors can differentially regulate this process.
HostingRepository	PRIDE
AnnounceDate	2024-01-30
AnnouncementXML	Submission_2024-01-30_02:47:42.216.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kelly Stecker
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF; Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2024-01-17 05:08:10	ID requested
⏵ 1	2024-01-30 02:47:42	announced

10.1038/S41556-024-01366-2;

submitter keyword: Protein Complex, CAMSAPs, Nucleation, trafficking,Microtubule, Tubulin

Kelly E. Stecker
contact affiliation	Utrecht University
contact email	k.e.stecker@uu.nl
lab head
Kelly Stecker
contact affiliation	Utrecht University
contact email	k.e.stecker@uu.nl
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/2024/01/PXD048637

PRIDE project URI

• PRIDE
  1. PXD048637
     1. Label: PRIDE project
     2. Name: CAMSAP-driven microtubule release from γ-TuRC and its regulation by nucleation-promoting factors