PXD048637-1

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

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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