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. 1998 Oct 15;17(20):5855–5867. doi: 10.1093/emboj/17.20.5855

KIF2beta, a new kinesin superfamily protein in non-neuronal cells, is associated with lysosomes and may be implicated in their centrifugal translocation.

N Santama 1, J Krijnse-Locker 1, G Griffiths 1, Y Noda 1, N Hirokawa 1, C G Dotti 1
PMCID: PMC1170913  PMID: 9774330

Abstract

Lysosomes concentrate juxtanuclearly in the region around the microtubule-organizing center by interaction with microtubules. Different experimental and physiological conditions can induce these organelles to move to the cell periphery by a mechanism implying a plus-end-directed microtubule-motor protein (a kinesin-like motor). The responsible kinesin-superfamily protein, however, is unknown. We have identified a new mouse isoform of the kinesin superfamily, KIF2beta, an alternatively spliced isoform of the known, neuronal kinesin, KIF2. Developmental expression pattern and cell-type analysis in vivo and in vitro reveal that KIF2beta is abundant at early developmental stages of the hippocampus but is then downregulated in differentiated neuronal cells, and it is mainly or uniquely expressed in non-neuronal cells while KIF2 remains exclusively neuronal. Electron microscopy of mouse fibroblasts and immunofluorescence of KIF2beta-transiently-transfected fibroblasts show KIF2 and KIF2beta primarily associated with lysosomes, and this association can be disrupted by detergent treatment. In KIF2beta-overexpressing cells, lysosomes (labeled with anti-lysosome-associated membrane protein-1) become abnormally large and peripherally located at some distance from their usual perinuclear positions. Overexpression of KIF2 or KIF2beta does not change the size or distribution of early, late and recycling endosomes nor does overexpression of different kinesin superfamily proteins result in changes in lysosome size or positioning. These results implicate KIF2beta as a motor responsible for the peripheral translocation of lysosomes.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aizawa H., Sekine Y., Takemura R., Zhang Z., Nangaku M., Hirokawa N. Kinesin family in murine central nervous system. J Cell Biol. 1992 Dec;119(5):1287–1296. doi: 10.1083/jcb.119.5.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andrews N. W. Lysosome recruitment during host cell invasion by Trypanosoma cruzi. Trends Cell Biol. 1995 Mar;5(3):133–137. doi: 10.1016/s0962-8924(00)88965-5. [DOI] [PubMed] [Google Scholar]
  3. Aniento F., Emans N., Griffiths G., Gruenberg J. Cytoplasmic dynein-dependent vesicular transport from early to late endosomes. J Cell Biol. 1993 Dec;123(6 Pt 1):1373–1387. doi: 10.1083/jcb.123.6.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bloom G. S., Endow S. A. Motor proteins 1: kinesins. Protein Profile. 1995;2(10):1105–1171. [PubMed] [Google Scholar]
  5. Bloom G. S., Endow S. A. Motor proteins. 1: kinesins. Protein Profile. 1994;1(10):1059–1116. [PubMed] [Google Scholar]
  6. Bottenstein J. E., Sato G. H. Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci U S A. 1979 Jan;76(1):514–517. doi: 10.1073/pnas.76.1.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burkhardt J. K., Echeverri C. J., Nilsson T., Vallee R. B. Overexpression of the dynamitin (p50) subunit of the dynactin complex disrupts dynein-dependent maintenance of membrane organelle distribution. J Cell Biol. 1997 Oct 20;139(2):469–484. doi: 10.1083/jcb.139.2.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chelsky D., Ralph R., Jonak G. Sequence requirements for synthetic peptide-mediated translocation to the nucleus. Mol Cell Biol. 1989 Jun;9(6):2487–2492. doi: 10.1128/mcb.9.6.2487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Debernardi S., Fontanella E., De Gregorio L., Pierotti M. A., Delia D. Identification of a novel human kinesin-related gene (HK2) by the cDNA differential display technique. Genomics. 1997 May 15;42(1):67–73. doi: 10.1006/geno.1997.4720. [DOI] [PubMed] [Google Scholar]
  10. Deng Y. P., Storrie B. Animal cell lysosomes rapidly exchange membrane proteins. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3860–3864. doi: 10.1073/pnas.85.11.3860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dillman J. F., 3rd, Dabney L. P., Pfister K. K. Cytoplasmic dynein is associated with slow axonal transport. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):141–144. doi: 10.1073/pnas.93.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dotti C. G., Sullivan C. A., Banker G. A. The establishment of polarity by hippocampal neurons in culture. J Neurosci. 1988 Apr;8(4):1454–1468. doi: 10.1523/JNEUROSCI.08-04-01454.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Feiguin F., Ferreira A., Kosik K. S., Caceres A. Kinesin-mediated organelle translocation revealed by specific cellular manipulations. J Cell Biol. 1994 Nov;127(4):1021–1039. doi: 10.1083/jcb.127.4.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ferris A. L., Brown J. C., Park R. D., Storrie B. Chinese hamster ovary cell lysosomes rapidly exchange contents. J Cell Biol. 1987 Dec;105(6 Pt 1):2703–2712. doi: 10.1083/jcb.105.6.2703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Geuze H. J., Stoorvogel W., Strous G. J., Slot J. W., Bleekemolen J. E., Mellman I. Sorting of mannose 6-phosphate receptors and lysosomal membrane proteins in endocytic vesicles. J Cell Biol. 1988 Dec;107(6 Pt 2):2491–2501. doi: 10.1083/jcb.107.6.2491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Griffiths G., Hoflack B., Simons K., Mellman I., Kornfeld S. The mannose 6-phosphate receptor and the biogenesis of lysosomes. Cell. 1988 Feb 12;52(3):329–341. doi: 10.1016/s0092-8674(88)80026-6. [DOI] [PubMed] [Google Scholar]
  18. Harada A., Takei Y., Kanai Y., Tanaka Y., Nonaka S., Hirokawa N. Golgi vesiculation and lysosome dispersion in cells lacking cytoplasmic dynein. J Cell Biol. 1998 Apr 6;141(1):51–59. doi: 10.1083/jcb.141.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Herman B., Albertini D. F. A time-lapse video image intensification analysis of cytoplasmic organelle movements during endosome translocation. J Cell Biol. 1984 Feb;98(2):565–576. doi: 10.1083/jcb.98.2.565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Heuser J. Changes in lysosome shape and distribution correlated with changes in cytoplasmic pH. J Cell Biol. 1989 Mar;108(3):855–864. doi: 10.1083/jcb.108.3.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hirokawa N. Kinesin and dynein superfamily proteins and the mechanism of organelle transport. Science. 1998 Jan 23;279(5350):519–526. doi: 10.1126/science.279.5350.519. [DOI] [PubMed] [Google Scholar]
  22. Hirokawa N. Organelle transport along microtubules - the role of KIFs. Trends Cell Biol. 1996 Apr;6(4):135–141. doi: 10.1016/0962-8924(96)10003-9. [DOI] [PubMed] [Google Scholar]
  23. Hirokawa N. The molecular mechanism of organelle transport along microtubules: the identification and characterization of KIFs (kinesin superfamily proteins). Cell Struct Funct. 1996 Oct;21(5):357–367. doi: 10.1247/csf.21.357. [DOI] [PubMed] [Google Scholar]
  24. Hollenbeck P. J., Swanson J. A. Radial extension of macrophage tubular lysosomes supported by kinesin. Nature. 1990 Aug 30;346(6287):864–866. doi: 10.1038/346864a0. [DOI] [PubMed] [Google Scholar]
  25. Hémar A., Olivo J. C., Williamson E., Saffrich R., Dotti C. G. Dendroaxonal transcytosis of transferrin in cultured hippocampal and sympathetic neurons. J Neurosci. 1997 Dec 1;17(23):9026–9034. doi: 10.1523/JNEUROSCI.17-23-09026.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. Lin S. X., Collins C. A. Immunolocalization of cytoplasmic dynein to lysosomes in cultured cells. J Cell Sci. 1992 Jan;101(Pt 1):125–137. doi: 10.1242/jcs.101.1.125. [DOI] [PubMed] [Google Scholar]
  28. Mattaj I. W., Englmeier L. Nucleocytoplasmic transport: the soluble phase. Annu Rev Biochem. 1998;67:265–306. doi: 10.1146/annurev.biochem.67.1.265. [DOI] [PubMed] [Google Scholar]
  29. Matteoni R., Kreis T. E. Translocation and clustering of endosomes and lysosomes depends on microtubules. J Cell Biol. 1987 Sep;105(3):1253–1265. doi: 10.1083/jcb.105.3.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. McGraw T. E., Dunn K. W., Maxfield F. R. Isolation of a temperature-sensitive variant Chinese hamster ovary cell line with a morphologically altered endocytic recycling compartment. J Cell Physiol. 1993 Jun;155(3):579–594. doi: 10.1002/jcp.1041550316. [DOI] [PubMed] [Google Scholar]
  31. Moore J. D., Endow S. A. Kinesin proteins: a phylum of motors for microtubule-based motility. Bioessays. 1996 Mar;18(3):207–219. doi: 10.1002/bies.950180308. [DOI] [PubMed] [Google Scholar]
  32. Morfini G., Quiroga S., Rosa A., Kosik K., Cáceres A. Suppression of KIF2 in PC12 cells alters the distribution of a growth cone nonsynaptic membrane receptor and inhibits neurite extension. J Cell Biol. 1997 Aug 11;138(3):657–669. doi: 10.1083/jcb.138.3.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mu F. T., Callaghan J. M., Steele-Mortimer O., Stenmark H., Parton R. G., Campbell P. L., McCluskey J., Yeo J. P., Tock E. P., Toh B. H. EEA1, an early endosome-associated protein. EEA1 is a conserved alpha-helical peripheral membrane protein flanked by cysteine "fingers" and contains a calmodulin-binding IQ motif. J Biol Chem. 1995 Jun 2;270(22):13503–13511. doi: 10.1074/jbc.270.22.13503. [DOI] [PubMed] [Google Scholar]
  34. Nakagawa T., Tanaka Y., Matsuoka E., Kondo S., Okada Y., Noda Y., Kanai Y., Hirokawa N. Identification and classification of 16 new kinesin superfamily (KIF) proteins in mouse genome. Proc Natl Acad Sci U S A. 1997 Sep 2;94(18):9654–9659. doi: 10.1073/pnas.94.18.9654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nakata T., Hirokawa N. Point mutation of adenosine triphosphate-binding motif generated rigor kinesin that selectively blocks anterograde lysosome membrane transport. J Cell Biol. 1995 Nov;131(4):1039–1053. doi: 10.1083/jcb.131.4.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nangaku M., Sato-Yoshitake R., Okada Y., Noda Y., Takemura R., Yamazaki H., Hirokawa N. KIF1B, a novel microtubule plus end-directed monomeric motor protein for transport of mitochondria. Cell. 1994 Dec 30;79(7):1209–1220. doi: 10.1016/0092-8674(94)90012-4. [DOI] [PubMed] [Google Scholar]
  37. Noda Y., Sato-Yoshitake R., Kondo S., Nangaku M., Hirokawa N. KIF2 is a new microtubule-based anterograde motor that transports membranous organelles distinct from those carried by kinesin heavy chain or KIF3A/B. J Cell Biol. 1995 Apr;129(1):157–167. doi: 10.1083/jcb.129.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Oda H., Stockert R. J., Collins C., Wang H., Novikoff P. M., Satir P., Wolkoff A. W. Interaction of the microtubule cytoskeleton with endocytic vesicles and cytoplasmic dynein in cultured rat hepatocytes. J Biol Chem. 1995 Jun 23;270(25):15242–15249. doi: 10.1074/jbc.270.25.15242. [DOI] [PubMed] [Google Scholar]
  39. Ogawa K., Mohri H. A dynein motor superfamily. Cell Struct Funct. 1996 Oct;21(5):343–349. doi: 10.1247/csf.21.343. [DOI] [PubMed] [Google Scholar]
  40. Okada Y., Yamazaki H., Sekine-Aizawa Y., Hirokawa N. The neuron-specific kinesin superfamily protein KIF1A is a unique monomeric motor for anterograde axonal transport of synaptic vesicle precursors. Cell. 1995 Jun 2;81(5):769–780. doi: 10.1016/0092-8674(95)90538-3. [DOI] [PubMed] [Google Scholar]
  41. Parton R. G., Dotti C. G., Bacallao R., Kurtz I., Simons K., Prydz K. pH-induced microtubule-dependent redistribution of late endosomes in neuronal and epithelial cells. J Cell Biol. 1991 Apr;113(2):261–274. doi: 10.1083/jcb.113.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rodríguez A., Samoff E., Rioult M. G., Chung A., Andrews N. W. Host cell invasion by trypanosomes requires lysosomes and microtubule/kinesin-mediated transport. J Cell Biol. 1996 Jul;134(2):349–362. doi: 10.1083/jcb.134.2.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Röttger S., White J., Wandall H. H., Olivo J. C., Stark A., Bennett E. P., Whitehouse C., Berger E. G., Clausen H., Nilsson T. Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus. J Cell Sci. 1998 Jan;111(Pt 1):45–60. doi: 10.1242/jcs.111.1.45. [DOI] [PubMed] [Google Scholar]
  44. Scholey J. M. Kinesin-II, a membrane traffic motor in axons, axonemes, and spindles. J Cell Biol. 1996 Apr;133(1):1–4. doi: 10.1083/jcb.133.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sekine Y., Okada Y., Noda Y., Kondo S., Aizawa H., Takemura R., Hirokawa N. A novel microtubule-based motor protein (KIF4) for organelle transports, whose expression is regulated developmentally. J Cell Biol. 1994 Oct;127(1):187–201. doi: 10.1083/jcb.127.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Slot J. W., Geuze H. J., Gigengack S., Lienhard G. E., James D. E. Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat. J Cell Biol. 1991 Apr;113(1):123–135. doi: 10.1083/jcb.113.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sperry A. O., Zhao L. P. Kinesin-related proteins in the mammalian testes: candidate motors for meiosis and morphogenesis. Mol Biol Cell. 1996 Feb;7(2):289–305. doi: 10.1091/mbc.7.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Storrie B., Desjardins M. The biogenesis of lysosomes: is it a kiss and run, continuous fusion and fission process? Bioessays. 1996 Nov;18(11):895–903. doi: 10.1002/bies.950181108. [DOI] [PubMed] [Google Scholar]
  49. Tardieux I., Webster P., Ravesloot J., Boron W., Lunn J. A., Heuser J. E., Andrews N. W. Lysosome recruitment and fusion are early events required for trypanosome invasion of mammalian cells. Cell. 1992 Dec 24;71(7):1117–1130. doi: 10.1016/s0092-8674(05)80061-3. [DOI] [PubMed] [Google Scholar]
  50. Toczyski D. P., Matera A. G., Ward D. C., Steitz J. A. The Epstein-Barr virus (EBV) small RNA EBER1 binds and relocalizes ribosomal protein L22 in EBV-infected human B lymphocytes. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3463–3467. doi: 10.1073/pnas.91.8.3463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Trayer I. P., John Smith K. Motoring down the highways of the cell. Trends Cell Biol. 1997 Jul;7(7):259–263. doi: 10.1016/S0962-8924(97)01083-0. [DOI] [PubMed] [Google Scholar]
  52. Vernos I., Raats J., Hirano T., Heasman J., Karsenti E., Wylie C. Xklp1, a chromosomal Xenopus kinesin-like protein essential for spindle organization and chromosome positioning. Cell. 1995 Apr 7;81(1):117–127. doi: 10.1016/0092-8674(95)90376-3. [DOI] [PubMed] [Google Scholar]
  53. Walczak C. E., Mitchison T. J., Desai A. XKCM1: a Xenopus kinesin-related protein that regulates microtubule dynamics during mitotic spindle assembly. Cell. 1996 Jan 12;84(1):37–47. doi: 10.1016/s0092-8674(00)80991-5. [DOI] [PubMed] [Google Scholar]
  54. Walczak C. E., Mitchison T. J. Kinesin-related proteins at mitotic spindle poles: function and regulation. Cell. 1996 Jun 28;85(7):943–946. doi: 10.1016/s0092-8674(00)81295-7. [DOI] [PubMed] [Google Scholar]
  55. Ward D. M., Leslie J. D., Kaplan J. Homotypic lysosome fusion in macrophages: analysis using an in vitro assay. J Cell Biol. 1997 Nov 3;139(3):665–673. doi: 10.1083/jcb.139.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wordeman L., Mitchison T. J. Identification and partial characterization of mitotic centromere-associated kinesin, a kinesin-related protein that associates with centromeres during mitosis. J Cell Biol. 1995 Jan;128(1-2):95–104. doi: 10.1083/jcb.128.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Yamazaki H., Nakata T., Okada Y., Hirokawa N. Cloning and characterization of KAP3: a novel kinesin superfamily-associated protein of KIF3A/3B. Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8443–8448. doi: 10.1073/pnas.93.16.8443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Yamazaki H., Nakata T., Okada Y., Hirokawa N. KIF3A/B: a heterodimeric kinesin superfamily protein that works as a microtubule plus end-directed motor for membrane organelle transport. J Cell Biol. 1995 Sep;130(6):1387–1399. doi: 10.1083/jcb.130.6.1387. [DOI] [PMC free article] [PubMed] [Google Scholar]

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