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Multiple domains of human CLASP contribute to microtubule dynamics and organization in vitro and in Xenopus egg extracts

Journal Article · · Cytoskeleton
DOI:https://doi.org/10.1002/cm.21005· OSTI ID:1623516
 [1];  [2];  [3]
  1. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology; DOE/OSTI
  2. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Science Division; Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst.
  3. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology
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Cytoplasmic linker associated proteins (CLASPs) comprise a class of microtubule (MT) plus end-binding proteins (1TIPs) that contribute to the dynamics and organization of MTs during many cellular processes, among them mitosis. Human CLASP proteins contain multiple MT-binding domains, including tumor overexpressed gene (TOG) domains, and a Ser-x-Ile-Pro (SxIP) motif known to target some 1TIPs though interaction with end-binding protein 1 (EB1). However, how individual domains contribute to CLASP function is poorly understood. We generated fulllength recombinant human CLASP1 and a series of truncation mutants and found that two N-terminal TOG domains make the strongest contribution to MT polymerization and bundling, but also identified a third TOG domain that further contributes to CLASP activity. Plus end tracking by CLASP requires the SxIP motif and interaction with EB1. The C-terminal coiled-coil domain mediates dimerization and association with many other factors, including the kinetochore motor centromere protein E (CENP-E), and the chromokinesin Xkid. Only the full-length protein was able to rescue spindle assembly in Xenopus egg extracts depleted of endogenous CLASP. Deletion of the C-terminal domain caused aberrant MT polymerization and dramatic spindle phenotypes, even with small amounts of added protein, indicating that proper localization of CLASP activity is essential to control MT polymerization during mitosis.
Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
National Institute of General Medical Sciences (NIGMS); USDOE Office of Science (SC)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1623516
Journal Information:
Cytoskeleton, Journal Name: Cytoskeleton Journal Issue: 3 Vol. 69; ISSN 1949-3584
Publisher:
WileyCopyright Statement
Country of Publication:
United States
Language:
English

References (38)

The Drosophila CLASP homologue, Mast/Orbit regulates the dynamic behaviour of interphase microtubules by promoting the pause state journal August 2007
Structural and mechanistic insights into microtubule end-binding proteins journal February 2010
The CENP-F-like Proteins HCP-1 and HCP-2 Target CLASP to Kinetochores to Mediate Chromosome Segregation journal April 2005
Role of CLASP2 in Microtubule Stabilization and the Regulation of Persistent Motility journal November 2006
Plus-End-Tracking Proteins and Their Interactions at Microtubule Ends journal June 2010
Stabilization of Overlapping Microtubules by Fission Yeast CLASP journal December 2007
CLASP Promotes Microtubule Rescue by Recruiting Tubulin Dimers to the Microtubule journal August 2010
Human CLASP1 Is an Outer Kinetochore Component that Regulates Spindle Microtubule Dynamics journal June 2003
Characterization of a Conserved “Threonine Clasp” in CAP-Gly Domains: Role of a Functionally Critical OH/π Interaction in Protein Recognition journal October 2008
The Ndc80 kinetochore complex forms oligomeric arrays along microtubules journal October 2010
CLASP fluxes its mitotic muscles journal January 2005
EB1 regulates microtubule dynamics and tubulin sheet closure in vitro journal March 2008
Golgi-derived CLASP-dependent microtubules control Golgi organization and polarized trafficking in motile cells journal August 2009
CLIPs and CLASPs and cellular dynamics journal June 2005
CLIP-170 facilitates the formation of kinetochore–microtubule attachments journal December 2005
The role of TOG domains in microtubule plus end dynamics journal September 2009
Centrosomes competent for parthenogenesis in Xenopus eggs support procentriole budding in cell-free extracts. journal November 1991
PRC1 Cooperates with CLASP1 to Organize Central Spindle Plasticity in Mitosis journal August 2009
Spatial regulation of CLASP affinity for microtubules by Rac1 and GSK3β in migrating epithelial cells journal June 2005
Xorbit/CLASP links dynamic microtubules to chromosomes in the Xenopus meiotic spindle journal January 2006
Clasp-mediated microtubule bundling regulates persistent motility and contact repulsion in Drosophila macrophages in vivo journal May 2010
XMAP215–EB1 Interaction Is Required for Proper Spindle Assembly and Chromosome Segregation in Xenopus Egg Extract journal June 2009
S. pombe CLASP needs dynein, not EB1 or CLIP170, to induce microtubule instability and slows polymerization rates at cell tips in a dynein-dependent manner journal September 2006
Mammalian CLASPs are required for mitotic spindle organization and kinetochore alignment journal August 2006
Orbit/Mast, the CLASP orthologue of Drosophila, is required for asymmetric stem cell and cystocyte divisions and development of the polarised microtubule network that interconnects oocyte and nurse cells during oogenesis journal March 2003
Phosphorylation of CLASP2 by GSK-3  regulates its interaction with IQGAP1, EB1 and microtubules journal July 2009
An EB1-binding motif acts as a microtubule tip localization signal text January 2009
Human CLASP1 Is an Outer Kinetochore Component that Regulates Spindle Microtubule Dynamics journal June 2003
Microtubule tip-interacting proteins: a view from both ends journal February 2011
XMAP215 Is a Processive Microtubule Polymerase journal January 2008
An EB1-Binding Motif Acts as a Microtubule Tip Localization Signal journal July 2009
Crystal Structure of a TOG Domain: Conserved Features of XMAP215/Dis1-Family TOG Domains and Implications for Tubulin Binding journal March 2007
Drosophila CLASP is required for the incorporation of microtubule subunits into fluxing kinetochore fibres journal December 2004
The role of TOG domains in microtubule plus end dynamics journal September 2009
PRC1 Cooperates with CLASP1 to Organize Central Spindle Plasticity in Mitosis journal August 2009
CLASP1 and CLASP2 bind to EB1 and regulate microtubule plus-end dynamics at the cell cortex journal January 2005
Measuring the Stoichiometry and Physical Interactions between Components Elucidates the Architecture of the Vertebrate Kinetochore journal October 2005
XMAP215–EB1 Interaction Is Required for Proper Spindle Assembly and Chromosome Segregation in Xenopus Egg Extract journal June 2009

Cited By (18)

Abelson phosphorylation of CLASP2 modulates its association with microtubules and actin journal March 2014
Facilitating identification of minimal protein binding domains by cross-linking mass spectrometry journal October 2017
Phosphorylation Dynamics Dominate the Regulated Proteome during Early Xenopus Development journal November 2017
Epiblast integrity requires CLASP and Dystroglycan-mediated microtubule anchoring to the basal cortex journal August 2013
TPX2 phosphorylation maintains metaphase spindle length by regulating microtubule flux journal August 2015
A mitotic SKAP isoform regulates spindle positioning at astral microtubule plus ends journal May 2016
CLASP2 binding to curved microtubule tips promotes flux and stabilizes kinetochore attachments journal November 2019
Xenopuscytoplasmic linker–associated protein 1 (XCLASP1) promotes axon elongation and advance of pioneer microtubules journal May 2013
RanGTP and CLASP1 cooperate to position the mitotic spindle journal August 2013
An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue journal June 2018
Human CLASP2 specifically regulates microtubule catastrophe and rescue journal May 2018
Crescerin uses a TOG domain array to regulate microtubules in the primary cilium text January 2015
Gene Expression Analysis in Ovarian Cancer – Faults and Hints from DNA Microarray Study journal January 2014
Regulators of spindle microtubules and their mechanisms: Living together matters: SPINDLE MICROTUBULES AND THEIR MECHANISMS journal January 2018
Developmentally Regulated GTP binding protein 1 (DRG1) controls microtubule dynamics journal August 2017
Insulin Induces Microtubule Stabilization and Regulates the Microtubule Plus-end Tracking Protein Network in Adipocytes journal July 2019
CLASP2 lattice-binding near microtubule plus ends stabilizes kinetochore attachments journal January 2019
Developmentally Regulated GTP binding protein 1 (DRG1) controls microtubule dynamics text January 2017

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Xenopus egg extract
cell biology
cytoplasmic linker associated proteins
microtubule dynamics
microtubule plus end-binding protein
mitotic spindle assembly