Properties of SEPT9 Isoforms and the requirement for GTP binding

C. Robertson, Stewart Church, H. Nagar, J. Price, Peter Hall, Hilary Russell

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Members of the evolutionarily conserved septin family of genes are emerging as key components of several cellular processes including membrane trafficking, cytokinesis, and cell-cycle control events. SEPT9 has been shown to have a complex genomic architecture, such that up to 15 different isoforms are possible by the shuffling of five alternate amino termini and three alternate carboxy termini. Genomic and transcriptional alterations of SEPT9 have been associated with neoplasia. The present study has used a Sept9-specific antibody to determine the pattern of isoform expression in a range of tumour cell lines. Western blot analysis indicated considerable variation in the relative amounts and isoform content of Sept9. Immunofluorescence studies showed a range of patterns of cytoplasmic localization ranging from mainly particulate to mainly filamentous. Expression constructs were also generated for each amino terminal isoform to investigate the patterns of localization of individual isoforms and the effects on cells of ectopic expression. The present study shows that the epsilon isoform appears filamentous in this overexpression system while the remaining isoforms are particulate and cytoplasmic. Transient transfection of individual constructs into tumour cell lines results in cell-cycle perturbation with a G2/M arrest and dramatic growth suppression, which was greatest in cell lines with the lowest amounts of endogenous Sept9. Similar phenotypic observations were made with GTP-binding mutants of all five N-terminal variants of Sept9. However, dramatic differences were observed in the kinetics of accumulation of wild-type versus mutant septin protein in transfected cells. In conclusion, the present study shows that the expression patterns of Sept9 protein are very varied in a panel of tumour cell lines and the functional studies are consistent with a model of septin function as a component of a molecular scaffold that contributes to diverse cellular functions. Alterations in the levels of Sept9 protein by overexpression of individual isoforms can clearly perturb cellular behaviour and may thus provide a mechanistic explanation for observations of deranged septin expression in neoplasia. Copyright © 2004 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Original languageEnglish
Pages (from-to)519-527
Number of pages9
JournalJournal of Pathology
Volume203
Issue number1
DOIs
Publication statusPublished - May 2004

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Guanosine Triphosphate
Protein Isoforms
Septins
Tumor Cell Line
Cytokinesis
Mutant Proteins
Cell Cycle Checkpoints
Ireland
Fluorescent Antibody Technique
Transfection
Neoplasms
Cell Cycle
Proteins
Western Blotting
Cell Line
Membranes
Antibodies
Growth
Genes

Cite this

Robertson, C., Church, S., Nagar, H., Price, J., Hall, P., & Russell, H. (2004). Properties of SEPT9 Isoforms and the requirement for GTP binding. Journal of Pathology, 203(1), 519-527. https://doi.org/10.1002/path.1551
Robertson, C. ; Church, Stewart ; Nagar, H. ; Price, J. ; Hall, Peter ; Russell, Hilary. / Properties of SEPT9 Isoforms and the requirement for GTP binding. In: Journal of Pathology. 2004 ; Vol. 203, No. 1. pp. 519-527.
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Robertson, C, Church, S, Nagar, H, Price, J, Hall, P & Russell, H 2004, 'Properties of SEPT9 Isoforms and the requirement for GTP binding', Journal of Pathology, vol. 203, no. 1, pp. 519-527. https://doi.org/10.1002/path.1551

Properties of SEPT9 Isoforms and the requirement for GTP binding. / Robertson, C.; Church, Stewart; Nagar, H.; Price, J.; Hall, Peter; Russell, Hilary.

In: Journal of Pathology, Vol. 203, No. 1, 05.2004, p. 519-527.

Research output: Contribution to journalArticle

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Robertson C, Church S, Nagar H, Price J, Hall P, Russell H. Properties of SEPT9 Isoforms and the requirement for GTP binding. Journal of Pathology. 2004 May;203(1):519-527. https://doi.org/10.1002/path.1551