Kinesin Motor Proteins and Neurodegeneration

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+ + + + Kinesin Motor+ Proteins and Neurodegeneration + + + P Related Publications + P Research Tools JUNE + + 2016 + + + + + + + v Meetings Kinesin Motor Proteins and Neurodegeneration

GRC - Signaling by Adhesion Kinesin motor proteins regulate mitosis and anterograde cargo Motors and Neurodegeneration: Cell Cycle Defects Receptors transport as exemplified by fast axonal transport (FAT) in neurons. News June 18-19.Lewiston, ME, US Neurons depend on kinesins for cell cycle regulation, especially the The neuropathological hallmarks of AD are beta amyloid (Ab) European Cytoskeleton assembly and function of the mitotic spindle, a macromolecular plaques and hyperphosphorylated tau neurofibrillary tangles. Forum - Cell Adhesion and structure composed primarily of microtubules (MTs) that undergo Ab is produced by enzymatic cleavage of amyloid precursor Migration cycles of polymerization and depolymerization to properly protein (APP) by beta-secretase 1 (BACE1) and the presenilin June 20-23. Cambridge, UK segregate duplicate chromosomes into separate daughter cells. (PS)-containing gamma-secretase complex2,3. In AD-associated In addition, the kinesin motors use MTs to transport cargo such as cell cycle defects, both Ab and tau inhibit Eg5 activity and its GRC - Muscle and Molecular Motors proteins, lipids, and mitochondria to the axon and axon terminal, interaction with MTs, which negatively affects mitotic spindle 9-11 July 17-22. West Dover, VT an essential process for normal neuron development, growth, structure and function (Fig. 1). A dysfunctional spindle results and communication as the cell body is the site of synthesis1. in the mis-segregation of chromosomes, aneuploidy/hyperploidy, GRC - Plant and Microbial and cell death9-11. Tau's inhibition of Eg5 requires excess levels of Cytoskeleton + 10

Aß / Tau Eg5 MT-bound tau . Neurons displaying aneuploidy/hyperploidy are Publications + August 14-19. Andover, NH + + + + increased in preclinical AD and selectively die as AD progresses. + + The Triangle Cytoskeleton Indeed, 90% of cell death in autopsied AD brains is comprised of 12 Meeting + + hyperploid neurons . September. NC, USA + + Eg5’s role in AD is not limited to cell cycle defects as mature, post- mitotic neurons express this protein. Eg5 mediates Ab-induced Cytoskeleton inhibition of long-term potentiation and loss of hippocampal + + Products chromosome spines/synapses13,14, at least partly through a reduction in + + + + cell surface trafficking of NMDA and nerve growth factor/p75 Actin Proteins metaphase 13 Activation Assays spindle neutrophin receptors . Antibodies Fig. 1. Kinesin motor Eg5 binding to MTs, essential for proper mitotic spindle structure and function, is inhibited by bA and tau. ECM Proteins PS1 Cargo Cargo ELISA Kits In neurodegenerative diseases such as Alzheimer’s disease (AD), GSK-3ß P G-LISA® Kits cell cycle defects (e.g., chromosome mis-segregation, abnormal P Pull-down Assays mitotic spindle structure/function, aneuploidy) and impaired PP1 Research Tools Tau Tau Motor Proteins FAT are pathophysiological hallmarks of AD, the most common Monomer Filaments Small G-Proteins form of dementia2,3. Indeed, FAT deficits and axonal swellings P P P P P occur before the classic neuropathological signs of dementia Tubulin & FtsZ Proteins P P (plaques and tangles), leading some researchers to posit that transport deficits are an early sign of neuron vulnerability to Contact Us Microtubule P: 1 (303) 322.2254 neurodegeneration4-8. This newsletter focuses on the role Eg5 F: 1 (303) 322.2257 (KSP, kinesin spindle protein, KIF11, kinesin-5) and kinesin-1 Anterograde Tau E: [email protected] have in AD-associated cell cycle and FAT defects, respectively. Eg5 is a kinesin motor that interacts with MTs and is essential W: cytoskeleton.com for mitotic spindle formation and function. Kinesin-1 isthe Fig. 2. Kinesin-1-mediated anterograde FAT is inhibited by GSK-3b-mediated prototypical kinesin that mediates FAT. For a discussion of the role phosphorylation of kinesin-1 light chains which causes dissociation of of the dynein motor in neurodegeneration, see the 2014 January/ kinesin-1 and its cargo. GSK-3b can be activated by PS1 and/or PP1 with February newsletter. the latter activated by hyper-phosphorylated tau filaments.

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Continued from Page 1 References Motors and Neurodegeneration: Axonal Transport Defects 1. Bass P.W. 1998. The role of motor proteins in establishing the microtubule arrays of axons and dendrites. J. Chem. Neuroanat. 14, 175-180. 2. Millecamps S. and Julien J.-P. 2013. Axonal transport deficits and neurodegenerative diseases. All 4 neurodegeneration-related proteins (APP, Ab, PS, and tau) inhibit kinesin-1- Nat. Rev. Neurosci. 14, 161-176. 15-17 3. Morfini G.A. et al. 2009. Minisymposium: Axonal transport defects in neurodegenerative mediated FAT. For APP and Ab disruption of FAT , various mechanisms have diseases. J. Neurosci. 29, 12776-12786. been reported, ranging from actin aggregation and dynamic changes in actin 4. Stokin G.B. et al. 2005. Axonopathy and transport deficits early in the pathogenesis of 16 Alzheimer’s disease. Science. 307, 1282-1288. polymerization to activation of casein kinase 2, which initiates a sequential cascade 5. Pigino G. et al. 2003. Alzheimer’s presenilin 1 mutations impair kinesin-based axonal transport. of kinesin-1 light chain phosphorylation and release of cargo from kinesin-117. J. Neurosci. 23, 4499-4508. 18 6. Lazarov O. et al. 2007. Impairments in fast axonal transport and motor neuron deficits in Interestingly, APP undergoes anterograde FAT via binding with kinesin-1 light transgenic mice expressing familial Alzheimer’s disease-linked mutant presenilin 1. J. Neurosci. chains as part of an axon membrane compartment which also contains BACE1 and 27, 7011-7020. 19-21 b 7. Ishihara T. et al. 1999. Age-dependent emergence and progression of a tauopathy in transgenic PS1 . In this way, APP could be cleaved into A during and/or after kinesin-1- mice overexpressing the shortest human tau isoform. Neuron. 24, 751-762. mediated FAT21. However, this finding has been refuted by others22. 8. Zhang B. et al. 2004. Retarded axonal transport of R406W mutant tau in transgenic mice with a neurodegenerative tauopathy. J. Neurosci. 24, 4657-4667. 9. Borysov S.I. et al. 2011. Alzheimer Ab disrupts the mitotic spindle and directly inhibits mitotic Both tau and mutant PS1 (M146V) regulate kinesin-1-mediated FAT via microtubule motors. Cell Cycle. 10, 1397-1410. dephosphorylation-stimulated activation of GSK-3b, resulting in GSK-3b-mediated 10. Bouge A.-L. and Parmentier M.-L. 2016. Tau excess impairs mitosis and kinesin-5 function, leading to aneuploidy and cell death. Dis. Model. Mech. 9, 307-319. phosphorylation of kinesin-1 light chains, which in turn induce separation of 11. Rossi G. et al. 2008. A new function of microtubule-associated protein tau: involvement in kinesin-1 and its cargo5 (Fig. 2). As a tau kinase, GSK-3b can mediate formation of chromosome stability. Cell Cycle. 7, 1788-1794. 12. Arendt T. et al. 2010. Selective cell death of hyperploid neurons in Alzheimer’s disease. Am J. pathological, hyper-phosphorylated tau, which dissociates from MTs and causes Pathol. 177, 15-20. MT depolymerization, followed by formation of tau filaments and eventually 13. Ari C. et al. 2014. Alzheimer Ab inhibition of eg5/kin5 reduces neurotrophin/transmitter 2,3 receptor function. Neurobiol. Aging. 35, 1839-1849. neurofibrillary tangles . The amino terminus of either tau filaments or C-terminal- 14. Freund R.K. et al. 2016. Inhibition of the motor protein Eg5/Kinesin-5 in amyloid b-mediated truncated, unbound monomers (C-terminus contains MT binding domains) inhibits impairment of hippocampal long-term potentiation and dendritic spine loss. Mol. Pharmacol. 89, 552-559. kinesin-dependent FAT. Here, GSK-3b activity is increased via protein phosphatase 15. Gunawardena S. and Goldstein L.S.B. 2001. Disruption of axonal transport and neuronal 1 (PP1)-mediated dephosphorylation-induced activation of GSK-3b23 (Fig. 2). These viability by amyloid precursor protein mutations in Drosophila. Neuron. 32, 389-401. 16. Hiruma H. et al. Glutamate and amyloid b-protein rapidly inhibit fast axonal transport in studies have led researchers to suggest that FAT is vulnerable to disruption early in cultured rat hippocampal neurons by different mechanisms. J. Neurosci. 23, 8967-8977. the pathophysiology of AD2-5; indeed, axonal/transport defects occur long before 17. Pigino G. et al. 2009. Disruption of fast axonal transport is a pathogenic mechanism for 4 intraneuronal amyloid beta. Proc. Natl. Acad. Sci. USA. 106, 5907-5912. amyloid deposition . Furthermore, depletion of kinesin-1 produces similar transport 18. Koo E.H. et al. 1990. Precursor of amyloid protein in Alzheimer disease undergoes fast deficits while also increasing levels of Ab peptide levels and deposits4. anterograde axonal transport. Proc. Natl. Acad. Sci. USA. 87, 1561-1565. 19. Ferreira A. et al. 1993. Intraneuronal compartments if the amyloid precursor protein. J. Neurosci. 13, 3112-3123. Conclusion 20. Kamal A. et al. 2000. Axonal transport of amyloid precursor protein is mediated by direct binding to the kinesin light chain subunit of kinesin-I. Neuron. 28, 449-459. 21. Kamal A. et al. 2001. Kinesin-mediated axonal transport of a membrane compartment Normal neuron physiology relies on kinesin motors for a variety of processes, containing b-secretase and presenilin-1 requires APP. Nature. 414, 643-648. including proper mitosis and FAT. Impaired kinesin function has a profound and 22. Lazarov O. et al. 2005. Axonal transport, amyloid precursor protein, kinesin-1, and the processing apparatus: Revisited. J. Neurosci. 25, 2386-2395. wide-spread effect on neuron health as kinesin motor dysfunctions are not only 23. LaPointe N.E. et al. 2009. The amino terminus of tau inhibits kinesin-dependent axonal observed in AD, but Huntington's disease24, upper and lower motor neuron diseases, transport: Implications for filament toxicity. J. Neurosci. Res. 87, 440-451. 2,3 24. Morfini G.A. et al. 2009. Pathogenic Huntingtin inhibits fast axonal transport by activating JNK3 and Charcot-Marie-Tooth peripheral neuropathy . To assist in studying how kinesin and phosphorylating kinesin. Nat. Neurosci. 12, 864-871. and dynein motor proteins regulate neuron health and survival, Cytoskeleton, Inc. offers purified kinesin motor proteins and dynein protein, along with kits to measure Kinesin & Dynein Proteins MT-induced motor ATPase activity. Product Source Purity Cat. # Amount CP01-A 2 x 25 µg CENP-E Motor Domain Protein H. sapiens >85% Kits and Assays CP01-XL 1 x 1 mg Product Cat. # Amount Chromokinesin Motor Domain Protein H. sapiens >85% CR01-A 2 x 25 µg ATPase ELIPA™ (enzyme-linked, colorimetric) Porcine BK051 96 assays Dynein (cytoplasmic) >80% CS-DN01 1 x 50 µg Kinetic quantitation of ATP hydrolysis (Kcat 0.05 to >1.0) brain CytoPhos™ Phosphate Assay (endpoint assay) EG01-A 2 x 25 µg BK054 1000 assays Eg5 Motor Domain Protein H. sapiens >85% Colorimetric assay for ATPases & GTPases (Kcat 0.01 to >1.0) EG01-B 10 x 25 µg GTPase ELIPA™ (enzyme-linked, colorimetric) BK052 96 assays KIFC3 Motor Domain Protein H. sapiens >85% KC01-A 2 x 25 µg Kinetic quantitation of GTP hydrolysis (Kcat 0.05 to >1.0) Kinesin ELIPA™ Biochem Kit BK060 96 assays KIF3C Motor Domain Protein H. sapiens >85% KF01-A 2 x 25 µg For real time kinetic and Vmax kinesin ATPase measurements Kinesin ATPase Endpoint Assay KIF7 motor domain H. sapiens >85% CS-KF51 1 x 100 µg BK053 1000 assays For endpoint measurement of kinesin ATPase activity Kinesin Heavy Chain Motor Domain KR01-A 2 x 25 µg H. sapiens >85% Protein KR01-XL 1 x 1 mg Microtubules MCAK Motor Domain Protein H. sapiens >85% MK01-A 2 x 25 µg MP01-A 2 x 25 µg MKLP1 Motor Domain Protein H. sapiens >85% Product Cat. # Amount MP01-XL 1 x 1 mg Microtubules, Pre-formed, lyophilized, MT002-A 4 x 500 µg porcine source, substrate for kinesin ATPase assays MT002-XL 1 x 10 mg MKLP2 Motor Domain Protein H. sapiens >85% CS-MP05 1 x 50 µg www.cytoskeleton.com