Home LiteratureArticle Details
PMID: 9813094 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Polymerization of purified yeast septins: evidence that organized filament arrays may not be required for septin function.

The Journal of cell biology ·Vol. 143 ·No. 3 ·1998-11-02 ·Pages 737-49

Frazier JA, Wong ML, Longtine MS, Pringle JR, Mann M, Mitchison TJ, Field C

Abstract

The septins are a family of proteins required for cytokinesis in a number of eukaryotic cell types. In budding yeast, these proteins are thought to be the structural components of a filament system present at the mother-bud neck, called the neck filaments. In this study, we report the isolation of a protein complex containing the yeast septins Cdc3p, Cdc10p, Cdc11p, and Cdc12p that is capable of forming long filaments in vitro. To investigate the relationship between these filaments and the neck filaments, we purified septin complexes from cells deleted for CDC10 or CDC11. These complexes were not capable of the polymerization exhibited by wild-type preparations, and analysis of the neck region by electron microscopy revealed that the cdc10Delta and cdc11Delta cells did not contain detectable neck filaments. These results strengthen the hypothesis that the septins are the major structural components of the neck filaments. Surprisingly, we found that septin dependent processes like cytokinesis and the localization of Bud4p to the neck still occurred in cdc10Delta cells. This suggests that the septins may be able to function in the absence of normal polymerization and the formation of a higher order filament structure.

MeSH Terms
Cell Cycle Proteins/genetics,isolation & purification,metabolism Cytoskeletal Proteins Fungal Proteins/genetics,isolation & purification,metabolism GTP Phosphohydrolases GTP-Binding Proteins/analysis Membrane Proteins Polymers Profilins Recombinant Fusion Proteins/genetics,isolation & purification,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Schizosaccharomyces pombe Proteins Transcription Factors
Chemicals
BUD4 protein, S cerevisiae CDC11 protein, S cerevisiae CDC12 protein, S cerevisiae CDC3 protein, S cerevisiae Cell Cycle Proteins Cytoskeletal Proteins Fungal Proteins Membrane Proteins Polymers Profilins Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Schizosaccharomyces pombe Proteins Transcription Factors cdc10 protein, S pombe CDC10 protein, S cerevisiae GTP Phosphohydrolases GTP-Binding Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Frazier J A
Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, California 94143, USA.
Wong M L
Longtine M S
Pringle J R
Mann M
Mitchison T J
Field C
References (44)
44 references, click to expand
  1. 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. 1991 Feb;112(4):535-44 PMID: 1993729
  2. The septins are required for the mitosis-specific activation of the Gin4 kinase.
    J Cell Biol. 1998 Nov 2;143(3):709-17 PMID: 9813092
  3. Localization and possible functions of Drosophila septins.
    Mol Biol Cell. 1995 Dec;6(12):1843-59 PMID: 8590810
  4. Cell division. Bud-site selection is only skin deep.
    Curr Biol. 1995 Nov 1;5(11):1213-5 PMID: 8574570
  5. In mouse brain profilin I and profilin II associate with regulators of the endocytic pathway and actin assembly.
    EMBO J. 1998 Feb 16;17(4):967-76 PMID: 9463375
  6. Genetic control of the cell division cycle in yeast. IV. Genes controlling bud emergence and cytokinesis.
    Exp Cell Res. 1971 Dec;69(2):265-76 PMID: 4950437
  7. Identification of a set of genes with developmentally down-regulated expression in the mouse brain.
    Biochem Biophys Res Commun. 1992 Jun 30;185(3):1155-61 PMID: 1378265
  8. Immunofluorescence methods for yeast.
    Methods Enzymol. 1991;194:565-602 PMID: 2005809
  9. The Drosophila peanut gene is required for cytokinesis and encodes a protein similar to yeast putative bud neck filament proteins.
    Cell. 1994 May 6;77(3):371-9 PMID: 8181057
  10. Members of the NAP/SET family of proteins interact specifically with B-type cyclins.
    J Cell Biol. 1995 Aug;130(3):661-73 PMID: 7622566
  11. Identification of a developmentally regulated septin and involvement of the septins in spore formation in Saccharomyces cerevisiae.
    J Cell Biol. 1996 Feb;132(3):399-411 PMID: 8636217
  12. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  13. A purified Drosophila septin complex forms filaments and exhibits GTPase activity.
    J Cell Biol. 1996 May;133(3):605-16 PMID: 8636235
  14. Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC11 gene product and the timing of events at the budding site.
    Dev Genet. 1991;12(4):281-92 PMID: 1934633
  15. Septins may form a ubiquitous family of cytoskeletal filaments.
    J Cell Biol. 1996 Sep;134(6):1345-8 PMID: 8830765
  16. Phenotypic analysis of temperature-sensitive yeast actin mutants.
    Cell. 1985 Feb;40(2):405-16 PMID: 3967297
  17. Guide to yeast genetics and molecular biology.
    Methods Enzymol. 1991;194:1-863 PMID: 2005781
  18. Yeast Saccharomyces cerevisiae selectable markers in pUC18 polylinkers.
    Yeast. 1990 Sep-Oct;6(5):363-6 PMID: 2220072
  19. Lymphocyte HEV adhesion variants differ in the expression of multiple gene sequences.
    Gene. 1990 Nov 15;95(2):279-84 PMID: 2174398
  20. Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases.
    Biochim Biophys Acta. 1966 Feb 7;112(2):346-62 PMID: 5329026
  21. Role of the yeast Gin4p protein kinase in septin assembly and the relationship between septin assembly and septin function.
    J Cell Biol. 1998 Nov 2;143(3):719-36 PMID: 9813093
  22. Nucleation of microtubule assembly by a gamma-tubulin-containing ring complex.
    Nature. 1995 Dec 7;378(6557):578-83 PMID: 8524390
  23. Evidence for kinesin-related proteins in the mitotic apparatus using peptide antibodies.
    J Cell Sci. 1992 Feb;101 ( Pt 2):303-13 PMID: 1629247
  24. A septin-based hierarchy of proteins required for localized deposition of chitin in the Saccharomyces cerevisiae cell wall.
    J Cell Biol. 1997 Oct 6;139(1):75-93 PMID: 9314530
  25. Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures.
    Genes Dev. 1997 Jun 15;11(12):1535-47 PMID: 9203580
  26. Subunit composition, protein interactions, and structures of the mammalian brain sec6/8 complex and septin filaments.
    Neuron. 1998 Jun;20(6):1111-22 PMID: 9655500
  27. Filament ring formation in the dimorphic yeast Candida albicans.
    J Cell Biol. 1983 Feb;96(2):486-93 PMID: 6339518
  28. Preparation of yeast cells for thin-section electron microscopy.
    Methods Enzymol. 1991;194:602-8 PMID: 2005811
  29. A Collection of cDNA Clones with Specific Expression Patterns in Mouse Brain.
    Eur J Neurosci. 1990;2(8):704-711 PMID: 12106288
  30. The GTPase superfamily: conserved structure and molecular mechanism.
    Nature. 1991 Jan 10;349(6305):117-27 PMID: 1898771
  31. Vectors for the inducible overexpression of glutathione S-transferase fusion proteins in yeast.
    Yeast. 1993 Jul;9(7):715-22 PMID: 8368005
  32. The BUD4 protein of yeast, required for axial budding, is localized to the mother/BUD neck in a cell cycle-dependent manner.
    J Cell Biol. 1996 Jul;134(2):413-27 PMID: 8707826
  33. A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae.
    Nucleic Acids Res. 1993 Jul 11;21(14):3329-30 PMID: 8341614
  34. Molecular cloning of a novel human cDNA homologous to CDC10 in Saccharomyces cerevisiae.
    Biochem Biophys Res Commun. 1994 Jul 15;202(1):82-7 PMID: 8037772
  35. SPR28, a sixth member of the septin gene family in Saccharomyces cerevisiae that is expressed specifically in sporulating cells.
    Microbiology. 1996 Oct;142 ( Pt 10):2897-905 PMID: 8885406
  36. A highly ordered ring of membrane-associated filaments in budding yeast.
    J Cell Biol. 1976 Jun;69(3):717-21 PMID: 773946
  37. Sequential assembly of myosin II, an IQGAP-like protein, and filamentous actin to a ring structure involved in budding yeast cytokinesis.
    J Cell Biol. 1998 Jan 26;140(2):355-66 PMID: 9442111
  38. ZDS1 and ZDS2, genes whose products may regulate Cdc42p in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Oct;16(10):5264-75 PMID: 8816439
  39. The septins: roles in cytokinesis and other processes.
    Curr Opin Cell Biol. 1996 Feb;8(1):106-19 PMID: 8791410
  40. Role of Bud3p in producing the axial budding pattern of yeast.
    J Cell Biol. 1995 May;129(3):767-78 PMID: 7730410
  41. A structural scaffolding of intermediate filaments in health and disease.
    Science. 1998 Jan 23;279(5350):514-9 PMID: 9438837
  42. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites.
    Gene. 1988 Dec 30;74(2):527-34 PMID: 3073106
  43. 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. 1987 Oct;7(10):3678-87 PMID: 3316985
  44. Components required for cytokinesis are important for bud site selection in yeast.
    J Cell Biol. 1993 Jul;122(2):373-86 PMID: 8320260
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.143.3.737
Published
1998-11-02
Pages
737-49
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2148147
Subset
IM
Grants
NIGMS NIH HHS · GM-31006 · United States
NIGMS NIH HHS · GM-23928 · United States
External Links
PubMed source DOI: 10.1083/jcb.143.3.737 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2