Skip to main content
SpringerLink
Log in

Expression of Nedd5, A Mammalian Septin, in Human Brain Tumors

  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

The septins are a family of cytoskeletal GTPases that play an essential role in cytokinesis in yeast and mammalian cells. Nedd5 is a mammalian septin known to associate with. actin-based structures such as the contractile ring and stress fibers. In the present study, we examined the expression of Nedd5 in a series of human brain tumor cell lines and surgical specimens by northern and western analyses. The temporal expression of Nedd5 in U373 astrocytoma cells at various timepoints throughout the cell cycle was determined by an analysis of lovastatin- and nocodazole-treated, synchronized cell populations. The intracellular localization of Nedd5 was determined by immunocytochemistry of steady state cultures and nocodazole-treated cultures enriched in M phase cells. The effects of inhibiting Nedd5 expression in human brain tumors was determined by stably transfecting U373 astrocytoma cells with an antisense-Nedd5 cDNA expression vector and by analyzing clones for Nedd5 expression by immunocytochemistry, morphological changes, cell growth and nuclear content. All human brain tumor cell lines and surgical specimens expressed Nedd5 transcript and protein. Synchronized U373 astrocytoma cells showed a relative increase in Nedd5 transcript levels from late G1 to G2M phases; and an increase in Nedd5 protein levels from S to G2M phases. Maximum expression of both transcript and protein levels was observed at the G2M phase. By immunocytochemistry, Nedd5 was concentrated at the cleavage furrow of mitotic cells. Double staining with Nedd5 and F-actin showed co-localization of Nedd5 with actin filaments except during cytokinesis. Antisense-Nedd5 expression led to an accumulation of nuclear content. These data suggest that Nedd5 is involved in the process of cytokinesis in human brain tumours. Nedd5 expression may be cell cycle-dependent with increased levels found at G2M phase. Blocking Nedd5 expression in astrocytoma cells by antisense interferes with the process of cytokinesis during cell division.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Hales KG, Bi E, Wu JQ, Adam JC, Yu IC, Pringle JR: Cytokinesis: an emerging unified theory for eukaryotes? Curr Opin Cell Biol 11: 717-725, 1999

    Google Scholar 

  2. Haarer BK, Pringle JR: Immunofluorescence localization of the Saccharomyces cerevisiae CDC12 gene product to the vicinity of the 10-nm filaments in the mother-bud neck. Mol Cell Biol 7: 3678-3687, 1987

    Google Scholar 

  3. Longtine MS, DeMarini DJ, Valencik ML, Al-Awar OS, Fares H, De Virgilio C, Pringle JR: The septins: roles in cytokinesis and other processes. Curr Opin Cell Biol 8: 106-119, 1996

    Google Scholar 

  4. Chant J: Septin scaffolds and cleavage planes in Saccharomyces. Cell 84: 187-190, 1996

    Google Scholar 

  5. Barral Y, Mermall V, Mooseker MS, Snyder M: Compartmentalization of the cell cortex by septins is required for maintenance of cell polarity in yeast. Mol Cell 5: 841-851, 2000

    Google Scholar 

  6. Takizawa PA, DeRisi JL, Wilhelm JE, Vale RD: Plasma membrane compartmentalization in yeast by messenger RNA transport and a septin diffusion barrier. Science 290: 341-344, 2000

    Google Scholar 

  7. Field CM, Kellogg D: Septins: cytoskeletal polymers or signalling GTPases?. Trends Cell Biol 9: 387-394, 1999

    Google Scholar 

  8. Neufeld TP, Rubin GM: The Drosophila peanut gene is required for cytokinesis and encodes a protein similar to yeast putative bud neck filament proteins. Cell 77: 371-379, 1994

    Google Scholar 

  9. Kinoshita M, Kumar S, Mizoguchi A, Ide C, Kinoshita A, Haraguchi T, Hiraoka Y, Noda M: Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures. Genes Dev 11: 1535-1547, 1997

    Google Scholar 

  10. Fares H, Goetsch L, Pringle JR: Identification of a developmentally regulated septin and involvement of the septins in spore formation in Saccharomyces cerevisiae. J Cell Biol 132: 399-411, 1996

    Google Scholar 

  11. Fares H, Peifer M, Pringle, JR: Localization and possible functions of Drosophila septins. Mol Biol Cell 6: 1843-1859, 1995

    Google Scholar 

  12. Hsu SC, Hazuka CD, Roth R, Foletti DL, Heuser J, Scheller RH: Subunit composition, protein interactions, and structures of the mammalian brain sec6/8 complex and septin filaments. Neuron 20: 1111-1122, 1998

    Google Scholar 

  13. Beites CL, Xie H, Bowser R, Trimble WS: The septin CDCrel-1 binds syntaxin and inhibits exocytosis. Nat Neurosci 2: 434-439, 1999

    Google Scholar 

  14. Osaka M, Rowley JD, Zeleznik-Le NJ: MSF (MLL septinlike fusion), a fusion partner gene of MLL, in a therapyrelated acute myeloid leukemia with a t(11; 17)(q23; q25). Proc Natl Acad Sci USA 96: 6428-6433, 1999

    Google Scholar 

  15. Kalikin LM, Sims HL, Petty EM: Genomic and expression analyses of alternatively spliced transcripts of the MLL septin-like fusion gene (MSF) that map to a 17q25 region of loss in breast and ovarian tumors Genomics 63: 165-172, 2000

    Google Scholar 

  16. McKie JM, Sutherland HF, Harvey E, Kim UJ, Scambler PJ: A human gene similar to Drosophila melanogaster peanut maps to the DiGeorge syndrome region of 22q11. Hum Genet 101: 6-12, 1997

    Google Scholar 

  17. Kinoshita A, Kinoshita M, Akiyama H, Tomimoto H, Akiguchi I, Kumar S, Noda M, Kimura J: Identification of septins in neurofibrillary tangles in Alzheimer's disease. Am J Pathol 153: 1551-1560, 1998

    Google Scholar 

  18. Larisch S, Yi Y, Lotan R, Kerner H, Eimerl S, Tony Parks W, Gottfried Y, Birkey Reffey S, de Caestecker MP, Danielpour D, Book-Melamed N, Timberg R, Duckett CS, Lechleider RJ, Steller H, Orly J, Kim SJ, Roberts AB: A novel mitochondrial septin-like protein, ARTS, mediates apoptosis dependent on its P-loop motif. Nat Cell Biol 2: 915-921, 2000

    Google Scholar 

  19. Rutka JT, Giblin JR, Hoifodt HK, Dougherty DV, Bell CW, McCulloch JR, Davis RL, Wilson CB, Rosenblum ML: Establishment and characterization of a cell line from a human gliosarcoma. Cancer Res 46: 5893-5902, 1986

    Google Scholar 

  20. Rutka JT, Giblin JR, Dougherty DV, Liu HC, McCulloch JR, Bell CW, Stern RS, Wilson CB, Rosenblum ML: Establishment and characterization of five cell lines derived from human malignant gliomas. Acta Neuropathol 75: 92-103, 1987

    Google Scholar 

  21. Dirks PB, Murakami M, Hubbard SL, Rutka JT: Cyclins and cyclin-dependent kinase expression in human astrocytoma cell lines. J Neuropathol Exp Neurol 56: 291-300, 1997

    Google Scholar 

  22. Xie H, Surka M, Howard J, Trimble WS: Characterization of the mammalian septin H5: distinct patterns of cytoskeletal and membrane association from other septin proteins. Cell Motil Cytoskeleton 43: 52-62, 1999

    Google Scholar 

  23. Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity”. Anal Biochem 137: 266-267, 1984 (Addendum)

    Google Scholar 

  24. Gossen M, Bujard H: Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci (USA) 89: 5547-5551, 1992

    Google Scholar 

  25. Kumar S, Tomooka Y, Noda M: Identification of a set of genes with developmentally down-regulated expression in the mouse brain. Biochem Biophys Res Commun 185: 1155-1161, 1992

    Google Scholar 

  26. Xue J, Wang X, Malladi CS, Kinoshita M, Milburn PJ, Lengyel I, Rostas JA, Robinson PJ: Phosphorylation of a new brain-specific septin, G-septin, by cGMP-dependent protein kinase. J Biol Chem 275: 10047-10056, 2000

    Google Scholar 

  27. Longtine MS, Theesfeld CL, McMillan JN, Weaver E, Pringle JR, Lew DJ: Septin-dependent assembly of a cell cycle-regulatory module in Saccharomyces cerevisiae. Mol Cell Biol 20: 4049-4061, 2000

    Google Scholar 

  28. Hartwell LH: Genetic control of the cell division cycle in yeast. IV. Genes controlling bud emergence and cytokinesis. Exp Cell Res 69: 265-276, 1971

    Google Scholar 

  29. Byers B, Goetsch L: Loss of filamentous ring in cytokinesis-defective mutants of budding yeast. J Cell Biol 70: 35, 1976

    Google Scholar 

  30. Field CM, al-Awar O, Rosenblatt J, Wong ML, Alberts B, Mitchison TJ: A purified Drosophila septin complex forms filaments and exhibits GTPase activity. J Cell Biol 133: 605-616, 1996

    Google Scholar 

  31. Kim HB, Haarer BK, Pringle JR: Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC3 gene product and the timing of events at the budding site. J Cell Biol 112: 535-544, 1991

    Google Scholar 

  32. Tsukita S, Hieda Y: A new 82-kD barbed end-capping protein (radixin) localized in the cell-to-cell adherens junction: purification and characterization. J Cell Biol 108: 2369-2382, 1989

    Google Scholar 

  33. Russell SE, McIlhatton MA, Burrows JF, Donaghy PG, Chanduloy S, Petty EM, Kalikin LM, Church SW, McIlroy S, Harkin DP, Keilty GW, Cranston AN, Weissenbach J, Hickey I, Johnston PG: Isolation and mapping of a human septin gene to a region on chromosome 17q, commonly deleted in sporadic epithelial ovarian tumors. Cancer Res 60: 4729-4734, 2000

    Google Scholar 

  34. Carroll CW, Altman R, Schieltz D, Yates JR, Kellogg D: The septins are required for the mitosis-specific activation of the Gin4 kinase. J Cell Biol 143: 709-717, 1998

    Google Scholar 

  35. Bayani J, Zielenska M, Marrano P, Kwan Ng Y, Taylor MD, Jay V, Rutka JT, Squire JA: Molecular cytogenetic analysis of medulloblastomas and supratentorial primitive neuroectodermal tumors by using conventional banding, comparative genomic hybridization, and spectral karyotyping. J Neurosurg 93: 437-448, 2000

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Division of Neurosurgery, The Arthur and Sonia Labatt Brain Tumour Research Centre, Toronto, Ontario, Canada

    Keiichi Sakai, Masanori Kurimoto, Atsushi Tsugu, Sherri L. Hubbard & William S. Trimble

  2. The Department of Cell Biology, The Hospital for Sick Children, The University of Toronto, Toronto, Ontario, Canada

    James T. Rutka

Authors
  1. Keiichi Sakai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Masanori Kurimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Atsushi Tsugu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sherri L. Hubbard
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. William S. Trimble
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. James T. Rutka
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sakai, K., Kurimoto, M., Tsugu, A. et al. Expression of Nedd5, A Mammalian Septin, in Human Brain Tumors. J Neurooncol 57, 169–177 (2002). https://doi.org/10.1023/A:1015721801075

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1015721801075

Search

Navigation