Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1997 Jun 2;16(11):3025–3034. doi: 10.1093/emboj/16.11.3025

Kinesin is essential for cell morphogenesis and polarized secretion in Neurospora crassa.

S Seiler 1, F E Nargang 1, G Steinberg 1, M Schliwa 1
PMCID: PMC1169921  PMID: 9214620

Abstract

Kinesin is a force-generating molecule that is thought to translocate organelles along microtubules, but its precise cellular function is still unclear. To determine the role of kinesin in vivo, we have generated a kinesin-deficient strain in the simple cell system Neurospora crassa. Null cells exhibit severe alterations in cell morphogenesis, notably hyphal extension, morphology and branching. Surprisingly, the movement of organelles visualized by video microscopy is hardly affected, but apical hyphae fail to establish a Spitzenkörper, an assemblage of secretory vesicles intimately linked to cell elongation and morphogenesis in Neurospora and other filamentous fungi. As cell morphogenesis depends on polarized secretion, our findings demonstrate that a step in the secretory pathway leading to cell shape determination and cell elongation cannot tolerate a loss of kinesin function. The defect is suggested to affect the transport of small, secretory vesicles to the site involved in protrusive activity, resulting in the uncoordinated insertion of new cell wall material over much of the cell surface. These observations have implications for the presumptive function of kinesin in more complex cell systems.

Full Text

The Full Text of this article is available as a PDF (462.9 KB).

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. Akins R. A., Lambowitz A. M. General method for cloning Neurospora crassa nuclear genes by complementation of mutants. Mol Cell Biol. 1985 Sep;5(9):2272–2278. doi: 10.1128/mcb.5.9.2272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Amaratunga A., Morin P. J., Kosik K. S., Fine R. E. Inhibition of kinesin synthesis and rapid anterograde axonal transport in vivo by an antisense oligonucleotide. J Biol Chem. 1993 Aug 15;268(23):17427–17430. [PubMed] [Google Scholar]
  4. Austin B., Hall R. M., Tyler B. M. Optimized vectors and selection for transformation of Neurospora crassa and Aspergillus nidulans to bleomycin and phleomycin resistance. Gene. 1990 Sep 1;93(1):157–162. doi: 10.1016/0378-1119(90)90152-h. [DOI] [PubMed] [Google Scholar]
  5. Bloom G. S., Endow S. A. Motor proteins 1: kinesins. Protein Profile. 1995;2(10):1105–1171. [PubMed] [Google Scholar]
  6. Brady S. T. A novel brain ATPase with properties expected for the fast axonal transport motor. Nature. 1985 Sep 5;317(6032):73–75. doi: 10.1038/317073a0. [DOI] [PubMed] [Google Scholar]
  7. Brady S. T., Pfister K. K., Bloom G. S. A monoclonal antibody against kinesin inhibits both anterograde and retrograde fast axonal transport in squid axoplasm. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1061–1065. doi: 10.1073/pnas.87.3.1061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Collinge A. J., Trinci A. P. Hyphal tips of wild-type and spreading colonial mutants of Neurospora crassa. Arch Microbiol. 1974;99(4):353–368. doi: 10.1007/BF00696249. [DOI] [PubMed] [Google Scholar]
  9. Dabora S. L., Sheetz M. P. The microtubule-dependent formation of a tubulovesicular network with characteristics of the ER from cultured cell extracts. Cell. 1988 Jul 1;54(1):27–35. doi: 10.1016/0092-8674(88)90176-6. [DOI] [PubMed] [Google Scholar]
  10. Elluru R. G., Bloom G. S., Brady S. T. Fast axonal transport of kinesin in the rat visual system: functionality of kinesin heavy chain isoforms. Mol Biol Cell. 1995 Jan;6(1):21–40. doi: 10.1091/mbc.6.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Euteneuer U., McIntosh J. R. Polarity of midbody and phragmoplast microtubules. J Cell Biol. 1980 Nov;87(2 Pt 1):509–515. doi: 10.1083/jcb.87.2.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fath K. R., Trimbur G. M., Burgess D. R. Molecular motors are differentially distributed on Golgi membranes from polarized epithelial cells. J Cell Biol. 1994 Aug;126(3):661–675. doi: 10.1083/jcb.126.3.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Ferreira A., Caceres A., Kosik K. S. Intraneuronal compartments of the amyloid precursor protein. J Neurosci. 1993 Jul;13(7):3112–3123. doi: 10.1523/JNEUROSCI.13-07-03112.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ferreira A., Niclas J., Vale R. D., Banker G., Kosik K. S. Suppression of kinesin expression in cultured hippocampal neurons using antisense oligonucleotides. J Cell Biol. 1992 May;117(3):595–606. doi: 10.1083/jcb.117.3.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hall D. H., Hedgecock E. M. Kinesin-related gene unc-104 is required for axonal transport of synaptic vesicles in C. elegans. Cell. 1991 May 31;65(5):837–847. doi: 10.1016/0092-8674(91)90391-b. [DOI] [PubMed] [Google Scholar]
  17. Harold F. M. To shape a cell: an inquiry into the causes of morphogenesis of microorganisms. Microbiol Rev. 1990 Dec;54(4):381–431. doi: 10.1128/mr.54.4.381-431.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Hirokawa N., Sato-Yoshitake R., Kobayashi N., Pfister K. K., Bloom G. S., Brady S. T. Kinesin associates with anterogradely transported membranous organelles in vivo. J Cell Biol. 1991 Jul;114(2):295–302. doi: 10.1083/jcb.114.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Hoyt M. A., He L., Loo K. K., Saunders W. S. Two Saccharomyces cerevisiae kinesin-related gene products required for mitotic spindle assembly. J Cell Biol. 1992 Jul;118(1):109–120. doi: 10.1083/jcb.118.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jung G., Wu X., Hammer J. A., 3rd Dictyostelium mutants lacking multiple classic myosin I isoforms reveal combinations of shared and distinct functions. J Cell Biol. 1996 Apr;133(2):305–323. doi: 10.1083/jcb.133.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Marks D. L., Larkin J. M., McNiven M. A. Association of kinesin with the Golgi apparatus in rat hepatocytes. J Cell Sci. 1994 Sep;107(Pt 9):2417–2426. doi: 10.1242/jcs.107.9.2417. [DOI] [PubMed] [Google Scholar]
  24. Oakley B. R., Rinehart J. E. Mitochondria and nuclei move by different mechanisms in Aspergillus nidulans. J Cell Biol. 1985 Dec;101(6):2392–2397. doi: 10.1083/jcb.101.6.2392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Patel N., Thierry-Mieg D., Mancillas J. R. Cloning by insertional mutagenesis of a cDNA encoding Caenorhabditis elegans kinesin heavy chain. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):9181–9185. doi: 10.1073/pnas.90.19.9181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pfister K. K., Wagner M. C., Stenoien D. L., Brady S. T., Bloom G. S. Monoclonal antibodies to kinesin heavy and light chains stain vesicle-like structures, but not microtubules, in cultured cells. J Cell Biol. 1989 Apr;108(4):1453–1463. doi: 10.1083/jcb.108.4.1453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Robb M. J., Wilson M. A., Vierula P. J. A fungal actin-related protein involved in nuclear migration. Mol Gen Genet. 1995 Jun 10;247(5):583–590. doi: 10.1007/BF00290350. [DOI] [PubMed] [Google Scholar]
  28. Saxton W. M., Hicks J., Goldstein L. S., Raff E. C. Kinesin heavy chain is essential for viability and neuromuscular functions in Drosophila, but mutants show no defects in mitosis. Cell. 1991 Mar 22;64(6):1093–1102. doi: 10.1016/0092-8674(91)90264-y. [DOI] [PubMed] [Google Scholar]
  29. Schleicher M., André B., Andréoli C., Eichinger L., Haugwitz M., Hofmann A., Karakesisoglou J., Stöckelhuber M., Noegel A. A. Structure/function studies on cytoskeletal proteins in Dictyostelium amoebae as a paradigm. FEBS Lett. 1995 Aug 1;369(1):38–42. doi: 10.1016/0014-5793(95)00579-x. [DOI] [PubMed] [Google Scholar]
  30. Schmitz F., Wallis K. T., Rho M., Drenckhahn D., Murphy D. B. Intracellular distribution of kinesin in chromaffin cells. Eur J Cell Biol. 1994 Feb;63(1):77–83. [PubMed] [Google Scholar]
  31. Scholey J. M., Porter M. E., Grissom P. M., McIntosh J. R. Identification of kinesin in sea urchin eggs, and evidence for its localization in the mitotic spindle. Nature. 1985 Dec 5;318(6045):483–486. doi: 10.1038/318483a0. [DOI] [PubMed] [Google Scholar]
  32. Sebald W., Neupert W., Weiss H. Preparation of Neurospora crassa mitochondria. Methods Enzymol. 1979;55:144–148. doi: 10.1016/0076-6879(79)55020-4. [DOI] [PubMed] [Google Scholar]
  33. Steinberg G., Schliwa M. Characterization of the biophysical and motility properties of kinesin from the fungus Neurospora crassa. J Biol Chem. 1996 Mar 29;271(13):7516–7521. doi: 10.1074/jbc.271.13.7516. [DOI] [PubMed] [Google Scholar]
  34. Steinberg G., Schliwa M. Organelle movements in the wild type and wall-less fz;sg;os-1 mutants of Neurospora crassa are mediated by cytoplasmic microtubules. J Cell Sci. 1993 Oct;106(Pt 2):555–564. doi: 10.1242/jcs.106.2.555. [DOI] [PubMed] [Google Scholar]
  35. Steinberg G., Schliwa M. The Neurospora organelle motor: a distant relative of conventional kinesin with unconventional properties. Mol Biol Cell. 1995 Nov;6(11):1605–1618. doi: 10.1091/mbc.6.11.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Steinhardt R. A., Bi G., Alderton J. M. Cell membrane resealing by a vesicular mechanism similar to neurotransmitter release. Science. 1994 Jan 21;263(5145):390–393. doi: 10.1126/science.7904084. [DOI] [PubMed] [Google Scholar]
  37. Tinsley J. H., Minke P. F., Bruno K. S., Plamann M. p150Glued, the largest subunit of the dynactin complex, is nonessential in Neurospora but required for nuclear distribution. Mol Biol Cell. 1996 May;7(5):731–742. doi: 10.1091/mbc.7.5.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vale R. D., Hotani H. Formation of membrane networks in vitro by kinesin-driven microtubule movement. J Cell Biol. 1988 Dec;107(6 Pt 1):2233–2241. doi: 10.1083/jcb.107.6.2233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Vale R. D., Reese T. S., Sheetz M. P. Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell. 1985 Aug;42(1):39–50. doi: 10.1016/s0092-8674(85)80099-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wright B. D., Henson J. H., Wedaman K. P., Willy P. J., Morand J. N., Scholey J. M. Subcellular localization and sequence of sea urchin kinesin heavy chain: evidence for its association with membranes in the mitotic apparatus and interphase cytoplasm. J Cell Biol. 1991 May;113(4):817–833. doi: 10.1083/jcb.113.4.817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Xiang X., Beckwith S. M., Morris N. R. Cytoplasmic dynein is involved in nuclear migration in Aspergillus nidulans. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2100–2104. doi: 10.1073/pnas.91.6.2100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Xiang X., Roghi C., Morris N. R. Characterization and localization of the cytoplasmic dynein heavy chain in Aspergillus nidulans. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9890–9894. doi: 10.1073/pnas.92.21.9890. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES