Analysis of Septin Reorganization at Cytokinesis Using Polarized Fluorescence Microscopy

Frontiers in Cell and Developmental Biology
Molly McQuilkenAmy S Gladfelter

Abstract

Septins are conserved filament-forming proteins that act in diverse cellular processes. They closely associate with membranes and, in some systems, components of the cytoskeleton. It is not well understood how filaments assemble into higher-order structuresin vivoor how they are remodeled throughout the cell cycle. In the budding yeastS. cerevisiae, septins are found through most of the cell cycle in an hourglass organization at the mother-bud neck until cytokinesis when the collar splits into two rings that disassemble prior to the next cell cycle. Experiments using polarized fluorescence microscopy have suggested that septins are arranged in ordered, paired filaments in the hourglass and undergo a coordinated 90° reorientation during splitting at cytokinesis. This apparent reorganization could be due to two orthogonal populations of filaments disassembling and reassembling or being preferentially retained at cytokinesis. In support of this idea, we report a decrease in septin concentration at the mother-bud neck during cytokinesis consistent with other reports and the timing of the decrease depends on known septin regulators including the Gin4 kinase. We took a candidate-based approach to examine what factors control reorient...Continue Reading

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Citations

Feb 6, 2018·Scientific Reports·Sara Orellana-MuñozCarlos R Vázquez de Aldana
Sep 11, 2019·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Talon ChandlerPatrick La Rivière
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Jul 7, 2018·Cytoskeleton·Joseph MarquardtErfei Bi
Mar 27, 2020·Frontiers in Cell and Developmental Biology·Carmen WiedemannCornelia A Deeg
Feb 10, 2021·G3 : Genes - Genomes - Genetics·Julie TakagiGregory C Finnigan
Jul 29, 2021·Molecular Biology of the Cell·Benjamin L WoodsAmy S Gladfelter

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Methods Mentioned

BETA
transmission electron microscopy
fluorescence microscopy
fluorescence imaging
PCR
polarization microscopy

Software Mentioned

FIJI
PolScope
MF
OpenPolScope
Excel
MATLAB

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