PXD011431

PXD011431 is an original dataset announced via ProteomeXchange.

Title	Insulin Induces Microtubule Stabilization and Regulates the Microtubule Plus-End Tracking Protein Network in Adipocytes
Description	Insulin-stimulated glucose uptake is known to involve microtubules, although the function of microtubules and the microtubule-regulating proteins involved in insulin action are poorly understood. CLASP2, a plus-end tracking microtubule-associated protein (+TIP) that controls microtubule dynamics, was recently implicated as the first +TIP associated with insulin-regulated glucose uptake. Here, using protein-specific targeted quantitative phosphoproteomics within 3T3-L1 adipocytes, we discovered that insulin regulates phosphorylation of the CLASP2 network members G2L1, MARK2, CLIP2, AGAP3 and CKAP5 as well as EB1, revealing the existence of a previously unknown microtubule-associated protein system that responds to insulin. To further investigate, G2L1 interactome studies within 3T3-L1 adipocytes revealed that G2L1 co-immunoprecipitates CLASP2 and CLIP2 as well as the master integrators of +TIP assembly, the end binding (EB) proteins. Live-cell total internal reflection fluorescence microscopy in adipocytes revealed G2L1 and CLASP2 colocalize on microtubule plus-ends. We found that while insulin increases the number of CLASP2-containing plus-ends, insulin treatment simultaneously decreases CLASP2-containing plus-end velocity. In addition, we discovered that insulin stimulates re-distribution of CLASP2 and G2L1 from exclusive plus-end tracking to “trailing” behind the growing tip of the microtubule. Insulin treatment increases -tubulin Lysine 40 acetylation, a mechanism that was observed to be regulated by a counterbalance between GSK3 and mTOR, and also led to microtubule stabilization. Our studies introduce insulin-stimulated microtubule stabilization and plus-end trailing of +TIPs as new modes of insulin action and reveal the likelihood that a network of microtubule-associated proteins synergize to coordinate insulin-regulated microtubule dynamics.
HostingRepository	PRIDE
AnnounceDate	2019-06-13
AnnouncementXML	Submission_2019-06-13_09:13:41.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD011431
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Paul Langlais
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Rattus norvegicus (Rat); NCBI TaxID: 10116;
ModificationList	Ammonia-loss; Phospho; Dehydrated; Oxidation; Acetyl
Instrument	Orbitrap Fusion Lumos; LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2018-10-22 00:49:16	ID requested
⏵ 1	2019-06-13 09:13:43	announced

Parker SS, Krantz J, Kwak EA, Barker NK, Deer CG, Lee NY, Mouneimne G, Langlais PR, Insulin Induces Microtubule Stabilization and Regulates the Microtubule Plus-end Tracking Protein Network in Adipocytes. Mol Cell Proteomics, 18(7):1363-1381(2019) [pubmed]

curator keyword: Biomedical

submitter keyword: CLASP2 G2L1 Insulin Microtubules

Paul R Langlais
contact affiliation	Department of Medicine/Division of Endocrinology, University of Arizona College of Medicine
contact email	langlais@deptofmed.arizona.edu
lab head
Paul Langlais
contact affiliation	University of Arizona
contact email	langlais@deptofmed.arizona.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD011431
     1. Label: PRIDE project
     2. Name: Insulin Induces Microtubule Stabilization and Regulates the Microtubule Plus-End Tracking Protein Network in Adipocytes