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

© The Author(s), 2011. This article is distributed under the terms of the Creative Commons Attribution 3.0 License. The definitive version was published in Journal of Cell Biology 193 (2011): 1065-1081, doi:10.1083/jcb.201012143.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.This work was supported by the National Science Foundation under grant No. MCB-0719126 to A.S. Gladfelter, the National Institute of Biomedical Imaging and Bioengineering under grant No. EB002583 to R. Oldenbourg, a Drexel CURE grant from the State of Pennsylvania Tobacco Settlement Fund, and National Institute of Neurological Disorders and Stroke grant NS48090- 06A to E.T. Spiliotis

Subunit-dependent modulation of septin assembly: Budding yeast septin Shs1 promotes ring and gauze formation

Substitution of specific terminal subunits within septin complexes and septin phosphorylation drive the formation of distinct higher-order septin assemblies in budding yeast

Cytokinesis in vertebrate cells initiates by contraction of an equatorial actomyosin network composed of randomly oriented filaments

© The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in eLife 6 (2017): e30867, doi:10.7554/eLife.30867.The actomyosin ring generates force to ingress the cytokinetic cleavage furrow in animal cells, yet its filament organization and the mechanism of contractility is not well understood. We quantified actin filament order in human cells using fluorescence polarization microscopy and found that cleavage furrow ingression initiates by contraction of an equatorial actin network with randomly oriented filaments. The network subsequently gradually reoriented actin filaments along the cell equator. This strictly depended on myosin II activity, suggesting local network reorganization by mechanical forces. Cortical laser microsurgery revealed that during cytokinesis progression, mechanical tension increased substantially along the direction of the cell equator, while the network contracted laterally along the pole-to-pole axis without a detectable increase in tension. Our data suggest that an asymmetric increase in cortical tension promotes filament reorientation along the cytokinetic cleavage furrow, which might have implications for diverse other biological processes involving actomyosin rings.DWG has received funding from the European Community’s Seventh Framework Programme FP7/2007-2013 under grant agreement no. 241548 (MitoSys) and no. 258068 (Systems Microscopy), an ERC Starting Grant under agreement no. 281198 (DIVIMAGE), and from the Austrian Science Fund (FWF) project no. SFB F34-06 (Chromosome Dynamics). FS has received funding from an EMBO long-term fellowship (ALTF 1447–2012). SM has received funding from Human Frontier Science Program cross-disciplinary fellowship (LT000096/2011)

SEPT9 occupies the terminal positions in septin octamers and mediates polymerization-dependent functions in abscission

Mammalian SEPT9 is positioned at the end of septin octamers and is necessary for octamer assembly into polymers necessary for abscission during cytokinesis

The chromosomal passenger complex and centralspindlin independently contribute to contractile ring assembly

In contrast to their sequential roles in midzone assembly, the CPC and centralspindlin act through independent mechanisms to regulate contractile ring assembly

Recruitment of the mitotic exit network to yeast centrosomes couples septin displacement to actomyosin constriction

The Mitotic Exit Network (MEN) promotes mitotic exit and cytokinesis but if and how MEN independently controls these two processes is unclear. Here, the authors report that MEN displaces septins from the cell division site to promote actomyosin ring constriction, independently of MEN control of mitotic exit