CoChat
Sign Up
Cited by 77Microtubule and mitosis dynamicsCellular transport and secretionNeuroscience and Neuropharmacology Research

A novel split kinesin assay identifies motor proteins that interact with distinct vesicle populations

Open access
Jenkins, Brian, Decker, Helena, Bentley, Marvin, Luisi, Julie, Banker, Gary·2012·Journal of Cell Biology
DOI 10.1083/jcb.201205070PMC PMC3514038
Open this paper in CoChat — free account

Abstract

Identifying the kinesin motors that interact with different vesicle populations is a longstanding and challenging problem with implications for many aspects of cell biology. Here we introduce a new live-cell assay to assess kinesin–vesicle interactions and use it to identify kinesins that bind to vesicles undergoing dendrite-selective transport in cultured hippocampal neurons. We prepared a library of “split kinesins,” comprising an axon-selective kinesin motor domain and a series of kinesin tail domains that can attach to their native vesicles; when the split kinesins were assembled by chemical dimerization, bound vesicles were misdirected into the axon. This method provided highly specific results, showing that three Kinesin-3 family members—KIF1A, KIF13A, and KIF13B—interacted with dendritic vesicle populations. This experimental paradigm allows a systematic approach to evaluate motor–vesicle interactions in living cells.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1083/jcb.201205070

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Kinesin-1 and kinesin-3 are major drivers of anterograde transport in axons, trafficking endosomes, lysosomes, mRNA, and other cargoes.

    “Kinesin-1 and kinesin-3 are major drivers of anterograde transport in axons, trafficking endosomes, lysosomes, mRNA, and other cargoes (Jenkins et al., 2012; Beaudet et al., 2024; Nagpal et al., 2024).”

    From Tau hyperphosphorylation impairs cooperative binding to microtubules and perturbs organelle trafficking in neurons (2026) · cites this paper as Jenkins et al., 2012

    The Discussion (page unavailable) describes KIF1A, a kinesin-3 motor, as a principal motor for axonal transport of synaptic vesicle proteins and notes that a kinesin-1 motor transports some axonal cargoes. The Introduction broadly includes RNA complexes among kinesin cargoes, but the paper does not establish that kinesin-1 and kinesin-3 are major axonal drivers of endosomes, lysosomes, and mRNA specifically. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

No references could be resolved yet.

Research works printed but not linked: 44

  1. Akhmanova A. Hammer J.A. III. 2010. Linking molecular motors to membrane cargo. Curr. Opin. Cell Biol. 22: 479– 487. 10.1016/j.ceb.2010.04.008 20466533 PMC3393125 · DOI
  2. Belshaw P.J. Ho S.N. Crabtree G.R. Schreiber S.L.. 1996. Controlling protein association and subcellular localization with a synthetic ligand that induces heterodimerization of proteins. Proc. Natl. Acad. Sci. USA. 93: 4604– 4607. 10.1073/pnas.93.10.4604 8643450 PMC39324 · DOI
  3. Burack M.A. Silverman M.A. Banker G.. 2000. The role of selective transport in neuronal protein sorting. Neuron. 26: 465– 472. 10.1016/S0896-6273(00)81178-2 10839364 · DOI
  4. Chu P.J. Rivera J.F. Arnold D.B.. 2006. A role for Kif17 in transport of Kv4.2. J. Biol. Chem. 281: 365– 373. 10.1074/jbc.M508897200 16257958 · DOI
  5. Friedman D.S. Vale R.D.. 1999. Single-molecule analysis of kinesin motility reveals regulation by the cargo-binding tail domain. Nat. Cell Biol. 1: 293– 297. 10.1038/13008 10559942 · DOI
  6. Gan Y. McGraw T.E. Rodriguez-Boulan E.. 2002. The epithelial-specific adaptor AP1B mediates post-endocytic recycling to the basolateral membrane. Nat. Cell Biol. 4: 605– 609. 12105417
  7. Guillaud L. Setou M. Hirokawa N.. 2003. KIF17 dynamics and regulation of NR2B trafficking in hippocampal neurons. J. Neurosci. 23: 131– 140. 12514209 PMC6742138
  8. Henthorn K.S. Roux M.S. Herrera C. Goldstein L.S.. 2011. A role for kinesin heavy chain in controlling vesicle transport into dendrites in Drosophila. Mol. Biol. Cell. 22: 4038– 4046. 10.1091/mbc.E10-07-0572 21880894 PMC3204066 · DOI
  9. Hirokawa N. Noda Y.. 2008. Intracellular transport and kinesin superfamily proteins, KIFs: structure, function, and dynamics. Physiol. Rev. 88: 1089– 1118. 10.1152/physrev.00023.2007 18626067 · DOI
  10. Hirokawa N. Niwa S. Tanaka Y.. 2010. Molecular motors in neurons: transport mechanisms and roles in brain function, development, and disease. Neuron. 68: 610– 638. 10.1016/j.neuron.2010.09.039 21092854 · DOI · Paper page
  11. Horiguchi K. Hanada T. Fukui Y. Chishti A.H.. 2006. Transport of PIP3 by GAKIN, a kinesin-3 family protein, regulates neuronal cell polarity. J. Cell Biol. 174: 425– 436. 10.1083/jcb.200604031 16864656 PMC2064238 · DOI
  12. Huang C.F. Banker G.. 2011. The translocation selectivity of the kinesins that mediate neuronal organelle transport. Traffic. 13: 549– 564. 10.1111/j.1600-0854.2011.01325.x PMC3967410 22212743 · DOI
  13. Jacobson C. Schnapp B. Banker G.A.. 2006. A change in the selective translocation of the Kinesin-1 motor domain marks the initial specification of the axon. Neuron. 49: 797– 804. 10.1016/j.neuron.2006.02.005 16543128 · DOI
  14. Jareb M. Banker G.. 1998. The polarized sorting of membrane proteins expressed in cultured hippocampal neurons using viral vectors. Neuron. 20: 855– 867. 10.1016/S0896-6273(00)80468-7 9620691 · DOI
  15. Kaech S. Banker G.. 2006. Culturing hippocampal neurons. Nat. Protoc. 1: 2406– 2415. 10.1038/nprot.2006.356 17406484 · DOI
  16. Kaech S. Huang C.F. Banker G.. 2012. Short-term high-resolution imaging of developing hippocampal neurons in culture. Cold Spring Harb Protoc. 2012: 340– 343. 22383653 PMC4438678
  17. Kapitein L.C. Schlager M.A. Kuijpers M. Wulf P.S. van Spronsen M. MacKintosh F.C. Hoogenraad C.C.. 2010. Mixed microtubules steer dynein-driven cargo transport into dendrites. Curr. Biol. 20: 290– 299. 10.1016/j.cub.2009.12.052 20137950 · DOI · Paper page
  18. Kennedy M.J. Davison I.G. Robinson C.G. Ehlers M.D.. 2010. Syntaxin-4 defines a domain for activity-dependent exocytosis in dendritic spines. Cell. 141: 524– 535. 10.1016/j.cell.2010.02.042 20434989 PMC2874581 · DOI · Paper page
  19. Lee J.R. Shin H. Ko J. Choi J. Lee H. Kim E.. 2003. Characterization of the movement of the kinesin motor KIF1A in living cultured neurons. J. Biol. Chem. 278: 2624– 2629. 10.1074/jbc.M211152200 12435738 · DOI
  20. Leterrier C. Vacher H. Fache M.P. d’Ortoli S.A. Castets F. Autillo-Touati A. Dargent B.. 2011. End-binding proteins EB3 and EB1 link microtubules to ankyrin G in the axon initial segment. Proc. Natl. Acad. Sci. USA. 108: 8826– 8831. 10.1073/pnas.1018671108 21551097 PMC3102358 · DOI
  21. Lewis T.L. Jr Mao T. Svoboda K. Arnold D.B.. 2009. Myosin-dependent targeting of transmembrane proteins to neuronal dendrites. Nat. Neurosci. 12: 568– 576. 10.1038/nn.2318 19377470 PMC2937175 · DOI
  22. Lo K.Y. Kuzmin A. Unger S.M. Petersen J.D. Silverman M.A.. 2011. KIF1A is the primary anterograde motor protein required for the axonal transport of dense-core vesicles in cultured hippocampal neurons. Neurosci. Lett. 491: 168– 173. 10.1016/j.neulet.2011.01.018 21256924 · DOI
  23. Lo Presti L. Martin S.G.. 2011. Shaping fission yeast cells by rerouting actin-based transport on microtubules. Curr. Biol. 21: 2064– 2069. 10.1016/j.cub.2011.10.033 22137473 · DOI
  24. Macaskill A.F. Rinholm J.E. Twelvetrees A.E. Arancibia-Carcamo I.L. Muir J. Fransson A. Aspenstrom P. Attwell D. Kittler J.T.. 2009. Miro1 is a calcium sensor for glutamate receptor-dependent localization of mitochondria at synapses. Neuron. 61: 541– 555. 10.1016/j.neuron.2009.01.030 19249275 PMC2670979 · DOI
  25. Maxfield F.R. McGraw T.E.. 2004. Endocytic recycling. Nat. Rev. Mol. Cell Biol. 5: 121– 132. 10.1038/nrm1315 15040445 · DOI
  26. May P. Woldt E. Matz R.L. Boucher P.. 2007. The LDL receptor-related protein (LRP) family: an old family of proteins with new physiological functions. Ann. Med. 39: 219– 228. 10.1080/07853890701214881 17457719 · DOI
  27. Muthuswamy S.K. Gilman M. Brugge J.S.. 1999. Controlled dimerization of ErbB receptors provides evidence for differential signaling by homo- and heterodimers. Mol. Cell. Biol. 19: 6845– 6857. 10490623 PMC84681
  28. Nakagawa T. Setou M. Seog D. Ogasawara K. Dohmae N. Takio K. Hirokawa N.. 2000. A novel motor, KIF13A, transports mannose-6-phosphate receptor to plasma membrane through direct interaction with AP-1 complex. Cell. 103: 569– 581. 10.1016/S0092-8674(00)00161-6 11106728 · DOI
  29. Nakata T. Hirokawa N.. 2003. Microtubules provide directional cues for polarized axonal transport through interaction with kinesin motor head. J. Cell Biol. 162: 1045– 1055. 10.1083/jcb.200302175 12975348 PMC2172855 · DOI
  30. Nakata T. Niwa S. Okada Y. Perez F. Hirokawa N.. 2011. Preferential binding of a kinesin-1 motor to GTP-tubulin-rich microtubules underlies polarized vesicle transport. J. Cell Biol. 194: 245– 255. 10.1083/jcb.201104034 21768290 PMC3144414 · DOI · Paper page
  31. Rasband M.N. 2010. The axon initial segment and the maintenance of neuronal polarity. Nat. Rev. Neurosci. 11: 552– 562. 10.1038/nrn2852 20631711 · DOI · Paper page
  32. Rumbaugh G. Sia G.M. Garner C.C. Huganir R.L.. 2003. Synapse-associated protein-97 isoform-specific regulation of surface AMPA receptors and synaptic function in cultured neurons. J. Neurosci. 23: 4567– 4576. 12805297 PMC6740810
  33. Setou M. Nakagawa T. Seog D.H. Hirokawa N.. 2000. Kinesin superfamily motor protein KIF17 and mLin-10 in NMDA receptor-containing vesicle transport. Science. 288: 1796– 1802. 10.1126/science.288.5472.1796 10846156 · DOI
  34. Setou M. Seog D.H. Tanaka Y. Kanai Y. Takei Y. Kawagishi M. Hirokawa N.. 2002. Glutamate-receptor-interacting protein GRIP1 directly steers kinesin to dendrites. Nature. 417: 83– 87. 10.1038/nature743 11986669 · DOI
  35. Shin H. Wyszynski M. Huh K.H. Valtschanoff J.G. Lee J.R. Ko J. Streuli M. Weinberg R.J. Sheng M. Kim E.. 2003. Association of the kinesin motor KIF1A with the multimodular protein liprin-alpha. J. Biol. Chem. 278: 11393– 11401. 10.1074/jbc.M211874200 12522103 · DOI
  36. Silverman M.A. Kaech S. Jareb M. Burack M.A. Vogt L. Sonderegger P. Banker G.. 2001. Sorting and directed transport of membrane proteins during development of hippocampal neurons in culture. Proc. Natl. Acad. Sci. USA. 98: 7051– 7057. 10.1073/pnas.111146198 11416186 PMC34621 · DOI
  37. Silverman M.A. Kaech S. Ramser E.M. Lu X. Lasarev M.R. Nagalla S. Banker G.. 2010. Expression of kinesin superfamily genes in cultured hippocampal neurons. Cytoskeleton (Hoboken). 67: 784– 795. 20862690 PMC2996887
  38. Song A.H. Wang D. Chen G. Li Y. Luo J. Duan S. Poo M.M.. 2009. A selective filter for cytoplasmic transport at the axon initial segment. Cell. 136: 1148– 1160. 10.1016/j.cell.2009.01.016 19268344 · DOI · Paper page
  39. Terada S. Hirokawa N.. 2000. Moving on to the cargo problem of microtubule-dependent motors in neurons. Curr. Opin. Neurobiol. 10: 566– 573. 10.1016/S0959-4388(00)00129-X 11084318 · DOI
  40. Verhey K.J. Meyer D. Deehan R. Blenis J. Schnapp B.J. Rapoport T.A. Margolis B.. 2001. Cargo of kinesin identified as JIP scaffolding proteins and associated signaling molecules. J. Cell Biol. 152: 959– 970. 10.1083/jcb.152.5.959 11238452 PMC2198804 · DOI · Paper page
  41. Verhey K.J. Kaul N. Soppina V.. 2011. Kinesin assembly and movement in cells. Annu Rev Biophys. 40: 267– 288. 10.1146/annurev-biophys-042910-155310 21332353 · DOI · Paper page
  42. Wang X. Schwarz T.L.. 2009. The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. Cell. 136: 163– 174. 10.1016/j.cell.2008.11.046 19135897 PMC2768392 · DOI · Paper page
  43. Yonekawa Y. Harada A. Okada Y. Funakoshi T. Kanai Y. Takei Y. Terada S. Noda T. Hirokawa N.. 1998. Defect in synaptic vesicle precursor transport and neuronal cell death in KIF1A motor protein-deficient mice. J. Cell Biol. 141: 431– 441. 10.1083/jcb.141.2.431 9548721 PMC2148442 · DOI
  44. Yoshimura Y. Terabayashi T. Miki H.. 2010. Par1b/MARK2 phosphorylates kinesin-like motor protein GAKIN/KIF13B to regulate axon formation. Mol. Cell. Biol. 30: 2206– 2219. 10.1128/MCB.01181-09 20194617 PMC2863582 · DOI

Cited by

Sorted by citations

Authors
Jenkins, B., Decker, H., Bentley, M., Luisi, J., & Banker, G.
Affiliations
Oregon Health & Science University (US)
Corresponding author
Banker, Gary
ORCID
Venue
Journal of Cell Biology
Publisher
Rockefeller University Press
Published
2012-08-20
Volume
198
Issue
4
Pages
749-761
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
DOI
10.1083/jcb.201205070
PMC
PMC3514038
OpenAlex
W2072031181
Cited by
77
References
0
Citations by year
2026: 3 · 2025: 5 · 2024: 5 · 2023: 3 · 2022: 7 · 2021: 6 · 2020: 10 · 2019: 7 · 2018: 4 · 2017: 5 · 2016: 7 · 2015: 7 · 2014: 3 · 2013: 3 · 2012: 2
FWCI
2.12
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion, cc-by-nc-sa)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Funders
  • National Institute of Mental Health (MH066179, R01MH066179)
  • National Institute of Neurological Disorders and Stroke (P30NS06180)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
kinesin, dendritic transport, axonal transport, KIF1A, KIF13A, KIF13B, kinesin-3, hippocampal neurons, live-cell assay
MeSH terms
Animals, Cells, Cultured, Cytological Techniques, Female, Hippocampus, Motor Neurons, Pregnancy, Kinesins, Molecular Motor Proteins, Protein Transport, Cytoplasmic Vesicles, Rats

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review