Neurofilament Heavy Chain Side Arm Phosphorylation Regulates Axonal Transport of Neurofilaments

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2003 May 14

PMID 12743103

Citations 76

Authors

Steven Ackerley

Paul Thornhill

Andrew J Grierson

Janet Brownlees

Brian H Anderton

P Nigel Leigh

Christopher E Shaw

Christopher C J Miller

Affiliations

Soon will be listed here.

Abstract

Neurofilaments possess side arms that comprise the carboxy-terminal domains of neurofilament middle and heavy chains (NFM and NFH); that of NFH is heavily phosphorylated in axons. Here, we demonstrate that phosphorylation of NFH side arms is a mechanism for regulating transport of neurofilaments through axons. Mutants in which known NFH phosphorylation sites were mutated to preclude phosphorylation or mimic permanent phosphorylation display altered rates of transport in a bulk transport assay. Similarly, application of roscovitine, an inhibitor of the NFH side arm kinase Cdk5/p35, accelerates neurofilament transport. Analyses of neurofilament movement in transfected living neurons demonstrated that a mutant mimicking permanent phosphorylation spent a higher proportion of time pausing than one that could not be phosphorylated. Thus, phosphorylation of NFH slows neurofilament transport, and this is due to increased pausing in neurofilament movement.

Citing Articles

The lemur tail kinase family in neuronal function and disfunction in neurodegenerative diseases.

Larose A, Miller C, Morotz G Cell Mol Life Sci. 2024; 81(1):447.

PMID: 39520508 PMC: 11550312. DOI: 10.1007/s00018-024-05480-0.

Progressive Optic Neuropathy in Hydrocephalic Ccdc13 Mutant Mice Caused by Impaired Axoplasmic Transport at the Optic Nerve Head.

Wu M, Zhao X, Peng S, Zhang X, Ru J, Xie L Invest Ophthalmol Vis Sci. 2024; 65(13):5.

PMID: 39499510 PMC: 11540025. DOI: 10.1167/iovs.65.13.5.

Stimulating VAPB-PTPIP51 ER-mitochondria tethering corrects FTD/ALS mutant TDP43 linked Ca and synaptic defects.

Markovinovic A, Martin-Guerrero S, Morotz G, Salam S, Gomez-Suaga P, Paillusson S Acta Neuropathol Commun. 2024; 12(1):32.

PMID: 38395965 PMC: 10885568. DOI: 10.1186/s40478-024-01742-x.

Neurofilaments in health and Charcot-Marie-Tooth disease.

Kotaich F, Caillol D, Bomont P Front Cell Dev Biol. 2024; 11:1275155.

PMID: 38164457 PMC: 10758125. DOI: 10.3389/fcell.2023.1275155.

Common mechanisms underlying axonal transport deficits in neurodegenerative diseases: a mini review.

Yang X, Ma Z, Lian P, Xu Y, Cao X Front Mol Neurosci. 2023; 16:1172197.

PMID: 37168679 PMC: 10164940. DOI: 10.3389/fnmol.2023.1172197.

References

Marszalek J, Williamson T, Lee M, Xu Z, Hoffman P, Becher M . Neurofilament subunit NF-H modulates axonal diameter by selectively slowing neurofilament transport. J Cell Biol. 1996; 135(3):711-24. PMC: 2121055. DOI: 10.1083/jcb.135.3.711. View

Yabe J, Pimenta A, Shea T . Kinesin-mediated transport of neurofilament protein oligomers in growing axons. J Cell Sci. 1999; 112 ( Pt 21):3799-814. DOI: 10.1242/jcs.112.21.3799. View

Wang L, Ho C, Sun D, Liem R, Brown A . Rapid movement of axonal neurofilaments interrupted by prolonged pauses. Nat Cell Biol. 2000; 2(3):137-41. DOI: 10.1038/35004008. View

Pant H, Veeranna , Grant P . Regulation of axonal neurofilament phosphorylation. Curr Top Cell Regul. 2000; 36:133-50. DOI: 10.1016/s0070-2137(01)80006-6. View

Ackerley S, Grierson A, Brownlees J, Thornhill P, Anderton B, Leigh P . Glutamate slows axonal transport of neurofilaments in transfected neurons. J Cell Biol. 2000; 150(1):165-76. PMC: 2185569. DOI: 10.1083/jcb.150.1.165. View

Roy S, Coffee P, Smith G, Liem R, Brady S, Black M . Neurofilaments are transported rapidly but intermittently in axons: implications for slow axonal transport. J Neurosci. 2000; 20(18):6849-61. PMC: 6772820. View

Shah J, Flanagan L, Janmey P, Leterrier J . Bidirectional translocation of neurofilaments along microtubules mediated in part by dynein/dynactin. Mol Biol Cell. 2000; 11(10):3495-508. PMC: 15009. DOI: 10.1091/mbc.11.10.3495. View

Eidenmuller J, Fath T, Hellwig A, Reed J, Sontag E, Brandt R . Structural and functional implications of tau hyperphosphorylation: information from phosphorylation-mimicking mutated tau proteins. Biochemistry. 2000; 39(43):13166-75. DOI: 10.1021/bi001290z. View

Prahlad V, Helfand B, Langford G, Vale R, Goldman R . Fast transport of neurofilament protein along microtubules in squid axoplasm. J Cell Sci. 2000; 113 ( Pt 22):3939-46. DOI: 10.1242/jcs.113.22.3939. View

10.

Sanchez I, Hassinger L, Sihag R, Cleveland D, Mohan P, Nixon R . Local control of neurofilament accumulation during radial growth of myelinating axons in vivo. Selective role of site-specific phosphorylation. J Cell Biol. 2000; 151(5):1013-24. PMC: 2174358. DOI: 10.1083/jcb.151.5.1013. View

11.

Yabe J, Chylinski T, Wang F, Pimenta A, Kattar S, Linsley M . Neurofilaments consist of distinct populations that can be distinguished by C-terminal phosphorylation, bundling, and axonal transport rate in growing axonal neurites. J Neurosci. 2001; 21(7):2195-205. PMC: 6762414. View

12.

Wang L, Brown A . Rapid intermittent movement of axonal neurofilaments observed by fluorescence photobleaching. Mol Biol Cell. 2001; 12(10):3257-67. PMC: 60171. DOI: 10.1091/mbc.12.10.3257. View

13.

Utton M, Connell J, Asuni A, van Slegtenhorst M, Hutton M, de Silva R . The slow axonal transport of the microtubule-associated protein tau and the transport rates of different isoforms and mutants in cultured neurons. J Neurosci. 2002; 22(15):6394-400. PMC: 6758152. DOI: 20026670. View

14.

Rao M, Garcia M, Miyazaki Y, Gotow T, Yuan A, Mattina S . Gene replacement in mice reveals that the heavily phosphorylated tail of neurofilament heavy subunit does not affect axonal caliber or the transit of cargoes in slow axonal transport. J Cell Biol. 2002; 158(4):681-93. PMC: 2174004. DOI: 10.1083/jcb.200202037. View

15.

Brownlees J, Ackerley S, Grierson A, Jacobsen N, Shea K, Anderton B . Charcot-Marie-Tooth disease neurofilament mutations disrupt neurofilament assembly and axonal transport. Hum Mol Genet. 2002; 11(23):2837-44. DOI: 10.1093/hmg/11.23.2837. View

16.

de Waegh S, Lee V, Brady S . Local modulation of neurofilament phosphorylation, axonal caliber, and slow axonal transport by myelinating Schwann cells. Cell. 1992; 68(3):451-63. DOI: 10.1016/0092-8674(92)90183-d. View

17.

Shetty K, Link W, Pant H . cdc2-like kinase from rat spinal cord specifically phosphorylates KSPXK motifs in neurofilament proteins: isolation and characterization. Proc Natl Acad Sci U S A. 1993; 90(14):6844-8. PMC: 47029. DOI: 10.1073/pnas.90.14.6844. View

18.

Elhanany E, Jaffe H, Link W, Sheeley D, Gainer H, Pant H . Identification of endogenously phosphorylated KSP sites in the high-molecular-weight rat neurofilament protein. J Neurochem. 1994; 63(6):2324-35. DOI: 10.1046/j.1471-4159.1994.63062324.x. View

19.

Collard J, Cote F, Julien J . Defective axonal transport in a transgenic mouse model of amyotrophic lateral sclerosis. Nature. 1995; 375(6526):61-4. DOI: 10.1038/375061a0. View

20.

Tu P, Elder G, Lazzarini R, Nelson D, Trojanowski J, Lee V . Overexpression of the human NFM subunit in transgenic mice modifies the level of endogenous NFL and the phosphorylation state of NFH subunits. J Cell Biol. 1995; 129(6):1629-40. PMC: 2291190. DOI: 10.1083/jcb.129.6.1629. View