Distinct Functions for Mammalian CLASP1 and -2 During Neurite and Axon Elongation

Frontiers in Cellular Neuroscience
C L SayasNiels Galjart

Abstract

Mammalian cytoplasmic linker associated protein 1 and -2 (CLASP1 and -2) are microtubule (MT) plus-end tracking proteins that selectively stabilize MTs at the edge of cells and that promote MT nucleation and growth at the Golgi, thereby sustaining cell polarity. In vitro analysis has shown that CLASPs are MT growth promoting factors. To date, a single CLASP1 isoform (called CLASP1α) has been described, whereas three CLASP2 isoforms are known (CLASP2α, -β, and -γ). Although CLASP2β/γ are enriched in neurons, suggesting isoform-specific functions, it has been proposed that during neurite outgrowth CLASP1 and -2 act in a redundant fashion by modulating MT dynamics downstream of glycogen synthase kinase 3 (GSK3). Here, we show that in differentiating N1E-115 neuroblastoma cells CLASP1 and CLASP2 differ in their accumulation at MT plus-ends and display different sensitivity to GSK3-mediated phosphorylation, and hence regulation. More specifically, western blot (WB) analysis suggests that pharmacological inhibition of GSK3 affects CLASP2 but not CLASP1 phosphorylation and fluorescence-based microscopy data show that GSK3 inhibition leads to an increase in the number of CLASP2-decorated MT ends, as well as to increased CLASP2 staining...Continue Reading

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Citations

Apr 26, 2020·Journal of Cell Science·Elizabeth J LawrenceLuke M Rice
Feb 16, 2021·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Luísa T FerreiraHelder Maiato
Jun 8, 2021·Circulation Research·Gerard A MarchalCarol Ann Remme
Sep 1, 2021·Trends in Endocrinology and Metabolism : TEM·Skylar R Batty, Paul R Langlais

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Methods Mentioned

BETA
dissection
transfection
Immunoprecipitation
Confocal Microscopy
PCR
co-IPs
immunoprecipitations
acetylation
Knockout
RNA-Seq

Software Mentioned

ImageJ
R
Tophat
Ensembl
Cufflinks
SPSS17
NeuronJ
CLASP

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