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Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals
Journal article   Open access   Peer reviewed

Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals

Bradley S DeMay, Xiaobo Bai, Louisa Howard, Patricia Occhipinti, Rebecca A Meseroll, Elias T Spiliotis, Rudolf Oldenbourg and Amy S Gladfelter
The Journal of cell biology, v 193(6), pp 1065-1081
13 Jun 2011
DOI: https://doi.org/10.1083/jcb.201012143
PMID: 21670216
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https://doi.org/10.1083/jcb.201012143View
Published, Version of Record (VoR) Open

Abstract

Cell Line Green Fluorescent Proteins - metabolism Septins - genetics Humans Protein Multimerization Yeasts - chemistry Green Fluorescent Proteins - genetics Cytoskeleton - ultrastructure Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - metabolism Microscopy, Fluorescence - instrumentation Animals Microscopy, Polarization - instrumentation Microscopy, Polarization - methods Microscopy, Fluorescence - methods Cytoskeleton - metabolism Recombinant Fusion Proteins - genetics Septins - ultrastructure Septins - metabolism Yeasts - cytology Septins - chemistry Yeasts - metabolism
The septins are conserved, GTP-binding proteins important for cytokinesis, membrane compartmentalization, and exocytosis. However, it is unknown how septins are arranged within higher-order structures in cells. To determine the organization of septins in live cells, we developed a polarized fluorescence microscopy system to monitor the orientation of GFP dipole moments with high spatial and temporal resolution. When GFP was fused to septins, the arrangement of GFP dipoles reflected the underlying septin organization. We demonstrated in a filamentous fungus, a budding yeast, and a mammalian epithelial cell line that septin proteins were organized in an identical highly ordered fashion. Fluorescence anisotropy measurements indicated that septin filaments organized into pairs within live cells, just as has been observed in vitro. Additional support for the formation of pairs came from the observation of paired filaments at the cortex of cells using electron microscopy. Furthermore, we found that highly ordered septin structures exchanged subunits and rapidly rearranged. We conclude that septins assemble into dynamic, paired filaments in vivo and that this organization is conserved from yeast to mammals.

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Domestic collaboration
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Cell Biology
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