GTPase domain driven dimerization of SEPT7 is dispensable for the critical role of septins in fibroblast cytokinesis

Scientific Reports
Megha AbbeyManoj B Menon

Abstract

Septin 7 (SEPT7) has been described to be essential for successful completion of cytokinesis in mouse fibroblasts, and Sept7-deficiency in fibroblasts constitutively results in multinucleated cells which stop proliferation. Using Sept7(flox/flox)fibroblasts we generated a cellular system, where the cytokinetic defects of Cre-mediated deletion of the Sept7 gene can be rescued by ectopically expressed doxycycline-inducible wild type SEPT7. Using this system, we analyzed the ability of SEPT7-mutants with alterations in their GTPase domain-dependent dimerization to prevent multinucleation and rescue proliferation. Although biochemical analysis of the mutants demonstrates differences in homo- and/or hetero-polymerization, in GTP-binding and/or GTPase activities, all analyzed mutants were able to rescue the cytokinesis phenotype of Sept7(flox/flox)fibroblasts associated with Cre-mediated deletion of endogenous Sept7. These findings indicate that the ability of septins to assemble into well-defined SEPT7-dimerization dependent native filaments is dispensable for cytokinesis in fibroblasts and opens the way to search for other mechanisms of the involvement of SEPT7 in cytokinesis.

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Citations

May 26, 2017·ELife·Andrew Weems, Michael McMurray
Mar 31, 2018·Journal of Cellular and Molecular Medicine·Xinlu WangShiwu Zhang
Nov 21, 2018·Cold Spring Harbor Molecular Case Studies·Christopher J ForlenzaNeerav Shukla
Nov 25, 2016·Frontiers in Cell and Developmental Biology·Dimitrios Angelis, Elias T Spiliotis
May 11, 2018·Cytoskeleton·Megha AbbeyManoj B Menon
Jul 24, 2020·World Journal of Gastrointestinal Oncology·Jie SunShi-Wu Zhang
May 19, 2021·NPJ Precision Oncology·Sipeng ShenDavid C Christiani

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Methods Mentioned

BETA
GTPase
X-ray
light microscopy
GTPases
fluorescence microscopy
affinity purification
gel-filtration
gel filtration
pulldown
nucleotide exchange

Software Mentioned

Image J

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